N-sulfonylaminocarbonyl containing compounds

ABSTRACT

Compounds having two reactive functional groups are described that can be used to provide a connector group between a substrate and an amine-containing material. The first reactive functional group can be used to provide attachment to a surface of a substrate. The second reactive functional group is a N-sulfonylaminocarbonyl group that can be reacted with an amine-containing material, particularly a primary aliphatic amine, to form a carbonylimino-containing connector group. The invention also provides articles and methods for immobilizing amine-containing materials to a substrate.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. Ser. No. 10/713,174, filed onNov. 14, 2003, now U.S. Pat No. 7,169,933, the disclosure of which isherein incorporated by reference.

FIELD OF THE INVENTION

This invention provides compounds that include both a substrate-reactivegroup and a N-sulfonylaminocarbonyl group. The invention also providesarticles and methods for immobilizing amine-containing materials to asubstrate.

BACKGROUND

Amine-containing materials such as amine-containing analytes, aminoacids, DNA fragments, RNA fragments, protein fragments, organelles, andimmunoglobins immobilized on the surface of a substrate can be used innumerous applications. For example, immobilized biological amines can beused for the medical diagnosis of a disease or genetic defect, forbiological separations, or for detection of various biomolecules.

The attachment of amine-containing materials to a substrate is oftenachieved through the use of a tethering compound. A tethering compoundusually has two reactive functional groups separated by a linking group.One of the functional groups provides a means for anchoring thetethering compound to a substrate by reacting with a complementaryfunctional group on the surface of the substrate. A second reactivefunctional group can be selected to react with an amine-containingmaterial. The second reactive functional group can be, for example, anactivated acyl derivative, such as an N-hydroxysuccinimide ester, or acyclic azlactone. An amine-containing material can react with theN-hydroxysuccinimide ester to form a carboxamide resulting in thedisplacement of an N-hydroxysuccinimide fragment. An amine-containingmaterial can react with the cyclic azlactone resulting in an opening ofthe ring structure.

Although tethering compounds that include a group such as anN-hydroxysuccinimide ester or a cyclic azlactone can be highly reactivewith primary amine-containing materials, such tethering compounds cansuffer from a number of disadvantages. Many of the reactions withbiological amines are conducted in dilute aqueous solutions. Under theseconditions, the N-hydroxysuccinimide ester functional group is known toundergo rapid hydrolysis. This competing reaction can cause incompleteor inefficient immobilization of the amine-containing materials on thesubstrate. While cyclic azlactone functional groups are more stable tohydrolysis, cyclic azlactone groups tend to be syntheticallyincompatible with many groups that could be used to attach the tetheringcompound to a substrate.

SUMMARY

Compounds are provided that can function as tethering compounds forimmobilizing an amine-containing material to a substrate. The compoundsinclude two types of reactive functional groups. The first type ofreactive functional group is a substrate-reactive group capable ofreacting with a complementary functional group on the surface of asubstrate resulting in the attachment of a tethering group to thesubstrate. The second type of reactive functional group is aN-sulfonylaminocarbonyl derivative that can react with anamine-containing material by a nucleophilic displacement reaction. Aconnector group is formed between the substrate and the amine-containingmaterial. Articles and methods for immobilizing amine-containingmaterials to a substrate are also provided.

One aspect of the invention provides compounds that can be attached to asubstrate and that can react with an amine-containing material. Thecompounds are of Formula I:

wherein

X¹ is a substrate-reactive functional group selected from a carboxy,halocarbonyl, halocarbonyloxy, cyano, hydroxy, mercapto, isocyanato,halosilyl, alkoxysilyl, acyloxysilyl, azido, aziridinyl, haloalkyl,tertiary amino, primary aromatic amino, secondary aromatic amino,disulfide, alkyl disulfide, benzotriazolyl, phosphono, phosphoroamido,phosphato, or ethylenically unsaturated group;

Y¹ is a single bond or a divalent group selected from an alkylene,heteroalkylene, arylene, carbonyl, carbonyloxy, carbonylimino, oxy,thio, —NR^(d)— where R^(d) is hydrogen or alkyl, or combinationsthereof;

Z¹ is an alkyl, aryl, or —(CO)R^(a) wherein R^(a) together with R¹ andgroups to which they are attached form a four to eight memberedheterocyclic or heterobicyclic group having a nitrogen heteroatom and asulfur heteroatom, wherein said heterocyclic or heterobicyclic group canbe fused to an optional aromatic group, optional saturated orunsaturated cyclic group, or optional saturated or unsaturated bicyclicgroup;

R¹ is an alkyl, fluoroalkyl, chloroalkyl, aryl, —NR^(b)R^(c) whereinR^(b) and R^(c) are each an alkyl group or taken together with thenitrogen atom to which they are attached form a four to eight memberedcyclic group, or R¹ taken together with R^(a) and the groups to whichthey are attached form the four to eight membered heterocyclic orheterobicyclic group that can be fused to the optional aromatic group,optional saturated or unsaturated cyclic group, or optional saturated orunsaturated bicyclic group; and

r is equal to 1 when X¹ is a monovalent group or equal to 2 when X¹ is adivalent group. The compound of Formula I can be unsubstituted orsubstituted with a halo, alkyl, alkoxy, or combinations thereof.

Another aspect of the invention provides articles that include atethering group attached to a substrate. The tethering group is thereaction product of the substrate-reactive group X¹ in compounds ofFormula I with a complementary functional group on the surface of thesubstrate to form an ionic bond, covalent bond, or a combinationthereof. The substrate-attached tethering group has aN-sulfonylaminocarbonyl group capable of reacting with anamine-containing material.

Yet another aspect of the invention provides a method of immobilizing anamine-containing material to a substrate. The method involves preparinga substrate-attached tethering group by reacting the substrate-reactivegroup X¹ in compounds of Formula I with a complementary functional groupon a substrate; and reacting a N-sulfonylaminocarbonyl group of thesubstrate-attached tethering group with an amine-containing material toform a connector group between the substrate and the amine-containingmaterial.

The invention also provides a multilayer substrate that includes apolymeric layer, a layer of diamond-like glass, and a layer ofdiamond-like carbon positioned between the polymeric layer and thediamond-like glass layer. A tethering group that includes aN-sulfonylaminocarbonyl group can be attached to the diamond-like glasslayer of the multilayer substrate.

The above summary of the present invention is not intended to describeeach disclosed embodiment or every implementation of the presentinvention. The Figures and the detailed description that follow moreparticularly exemplify these embodiments.

BRIEF DESCRIPTION OF THE DRAWINGS

The above aspects may be more completely understood in consideration ofthe following detailed description of various embodiments in connectionwith the accompanying drawings, in which:

FIG. 1 is a confocal micrograph of various concentrations (50micrograms/ml, 25 micrograms/ml, 12.5 micrograms/ml, and 6.25micrograms/ml from left to right) of fluorescence labeled mouse IgGimmobilized by reacting with N-sulfonylaminocarbonyl tethering groupsattached to a substrate.

FIG. 2 is a confocal micrograph showing the capture of Staphylococcusaureus with IgG immobilized on a multilayer substrate of diamond-likeglass/diamond-like carbon/polyimide/diamond-like carbon/diamond-likeglass.

FIG. 3 is a confocal micrograph showing the exposure of Staphylococcusaureus with a multilayer substrate of diamond-like glass/diamond-likecarbon/polyimide/diamond-like carbon/diamond like glass without IgGimmobilized to the substrate with a connector group derived from acompound of Formula I.

FIG. 4 is a confocal micrograph showing capture of Staphylococcus aureuswith IgG immobilized on a multilayer substrate ofpolyimide/titanium/gold.

FIG. 5 is a confocal micrograph showing the exposure of Staphylococcusaureus to a multilayer substrate of polyimide/titanium/gold without IgGimmobilized to the substrate.

While the invention is amenable to various modifications and alternativeforms, specifics thereof have been shown by way of example in thedrawings and will be described in detail. It should be understood,however, that the intention is not to limit the invention to theparticular embodiments described. To the contrary, the intention is tocover all modifications, equivalents, and alternatives falling withinthe spirit and scope of the present disclosure.

DETAILED DESCRIPTION

Compounds having two reactive functional groups are described that canbe used to provide a connector group between a substrate and anamine-containing material. The first reactive functional group can beused to provide attachment of a tethering group to a surface of asubstrate. The second reactive functional group is aN-sulfonylaminocarbonyl group that can be reacted with anamine-containing material, particularly a primary aliphaticamine-containing material, to form a carbonylimino-containing connectorgroup. The invention also provides articles and methods for immobilizingamine-containing materials to a substrate.

Definitions

As used herein, the terms “a”, “an”, and “the” are used interchangeablywith “at least one” to mean one or more of the elements being described.

As used herein, the term “acyl” refers to a monovalent group of formula—(CO)R where R is an alkyl group and where (CO) used herein indicatesthat the carbon is attached to the oxygen with a double bond.

As used herein, the term “acyloxy” refers to a monovalent group offormula —O(CO)R where R is an alkyl group.

As used herein, the term “acyloxysilyl” refers to a monovalent grouphaving an acyloxy group attached to a Si (i.e., Si—O(CO)R where R is analkyl). For example, an acyloxysilyl can have a formula—Si[O(CO)R]_(3-n)L_(n) where n is an integer of 0 to 2 and L is ahalogen or alkoxy. Specific examples include —Si[O(CO)CH₃]₃,—Si[O(CO)CH₃]₂Cl, or —Si[O(CO)CH₃]Cl₂.

As used herein, the term “alkoxy” refers to a monovalent group offormula —OR where R is an alkyl group.

As used herein, the term “alkoxycarbonyl” refers to a monovalent groupof formula —(CO)OR where R is an alkyl group.

As used herein, the term “alkoxysilyl” refers to a group having analkoxy group attached to a Si (i.e., Si—OR where R is an alkyl). Forexample, an alkoxysilyl can have a formula —Si(OR)_(3-n)(L^(a))_(n)where n is an integer of 0 to 2 and L^(a) is a halogen or acyloxy.Specific examples include —Si(OCH₃)₃, —Si(OCH₃)₂Cl, or —Si(OCH₃)Cl₂.

As used herein, the term “alkyl” refers to a monovalent radical of analkane and includes groups that are linear, branched, cyclic, orcombinations thereof. The alkyl group typically has 1 to 30 carbonatoms. In some embodiments, the alkyl group contains 1 to 20 carbonatoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbonatoms. Examples of alkyl groups include, but are not limited to, methyl,ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl,n-hexyl, cyclohexyl, n-heptyl, n-octyl, and ethylhexyl.

As used herein, the term “alkyl disulfide” refers to a monovalent groupof formula —SSR where R is an alkyl group.

As used herein, the term “alkylene” refers to a divalent radical of analkane. The alkylene can be straight-chained, branched, cyclic, orcombinations thereof. The alkylene typically has 1 to 200 carbon atoms.In some embodiments, the alkylene contains 1 to 100, 1 to 80, 1 to 50, 1to 30, 1 to 20, 1 to 10, or 1 to 4 carbon atoms. The radical centers ofthe alkylene can be on the same carbon atom (i.e., an alkylidene) or ondifferent carbon atoms.

As used herein, the term “aralkyl” refers to a monovalent radical of thecompound R—Ar where Ar is an aromatic carbocyclic group and R is analkyl group.

As used herein, the term “aralkylene” refers to a divalent radical offormula —R—Ar— where Ar is an arylene group and R is an alkylene group.

As used herein, the term “aryl” refers to a monovalent aromaticcarbocyclic radical. The aryl can have one aromatic ring or can includeup to 5 carbocyclic ring structures that are connected to or fused tothe aromatic ring. The other ring structures can be aromatic,non-aromatic, or combinations thereof. Examples of aryl groups include,but are not limited to, phenyl, biphenyl, terphenyl, anthryl, naphthyl,acenaphthyl, anthraquinonyl, phenanthryl, anthracenyl, pyrenyl,perylenyl, and fluorenyl.

As used herein, the term “arylene” refers to a divalent radical of acarbocyclic aromatic compound having one to 5 rings that are connected,fused, or combinations thereof. In some embodiments, the arylene grouphas up to 5 rings, up to 4 rings, up to 3 rings, up to 2 rings, or onearomatic ring. For example, the arylene group can be phenylene.

As used herein, the term “azido” refers to a group of formula —N₃.

As used herein, the term “aziridinyl” refers to a cyclic monovalentradical of aziridine having the formula

where R^(d) is hydrogen or alkyl.

As used herein, the term “benzotriazolyl” refers to a monovalent grouphaving a benzene group fused to a triazolyl group. The formula for abenzotriazolyl group is C₆H₄N₃—.

As used herein, the term “carbonyl” refers to a divalent group offormula —(CO)—.

As used herein, the term “carbonylimino” refers to a divalent group offormula —(CO)NR⁴— where R⁴ is hydrogen, alkyl, or aryl.

As used herein, the term “carbonyloxy” refers to a divalent group offormula —(CO)O—.

As used herein, the term “carbonyloxycarbonyl” refers to a divalentgroup of formula —(CO)O(CO)—. Such a group is part of an anhydridecompound.

As used herein, the term “carbonylthio” refers to a divalent group offormula —(CO)S—.

As used herein, the term “carboxy” refers to a monovalent group offormula —(CO)OH.

As used herein, the term “chloroalkyl” refers to an alkyl having atleast one hydrogen atom replaced with a chlorine atom.

As used herein, the term “cyano” refers to a group of formula —CN.

As used herein, the term “disulfide” refers to a divalent group offormula —S—S—.

As used herein, the term “ethylenically unsaturated” refers to amonovalent group having a carbon-carbon double bond of formula —CY═CH₂where Y is hydrogen, alkyl, or aryl.

As used herein, the term “fluoroalkyl” refers to an alkyl having atleast one hydrogen atom replaced with a fluorine atom.

As used herein, the term “haloalkyl” refers to an alkyl having at leastone hydrogen atom replaced with a halogen selected from F, Cl, Br, or I.Perfluoroalkyl groups are a subset of haloalkyl groups.

As used herein, the term “halocarbonyloxy” refers to a monovalent groupof formula —O(CO)X where X is a halogen atom selected from F, Cl, Br, orI.

As used herein, the term “halocarbonyl” refers to a monovalent group offormula —(CO)X where X is a halogen atom selected from F, Cl, Br, or I.

As used herein, the term “halosilyl” refers to a group having a Siattached to a halogen (i.e., Si—X where X is a halogen). For example,the halosilyl group can be of formula —SiX_(3-n)(L^(b))_(n) where n isan integer of 0 to 2 and L^(b) is selected from an alkoxy, or acyloxy.Some specific examples include the groups —SiCl₃, —SiCl₂OCH₃, and—SiCl(OCH₃)₂.

As used herein, the term “heteroalkylene” refers to a divalent alkylenehaving one or more carbon atoms replaced with a sulfur, oxygen, orNR^(d) where R^(d) is hydrogen or alkyl. The heteroalkylene can belinear, branched, cyclic, or combinations thereof and can include up to400 carbon atoms and up to 30 heteroatoms. In some embodiments, theheteroalkylene includes up to 300 carbon atoms, up to 200 carbon atoms,up to 100 carbon atoms, up to 50 carbon atoms, up to 30 carbon atoms, upto 20 carbon atoms, or up to 10 carbon atoms.

As used herein, the term “hydroxy” refers to a group of formula —OH.

As used herein, the term “isocyanato” refers to a group of formula —NCO.

As used herein, the term “mercapto” refers to a group of formula —SH.

As used herein, the term “N-sulfonylaminocarbonyl” refers to a divalententity of formula —SO₂NZ^(a)(CO)— where Z^(a) is an alkyl, aryl, or partof a group structure.

As used herein, the term “oxy” refers to a divalent group of formula—O—.

As used herein, the term “oxycarbonylimino” refers to a divalent groupof formula —O(CO)NR⁴— where R⁴ is hydrogen, alkyl, or aryl.

As used herein, the term “oxycarbonyloxy” refers to a divalent group offormula —O(CO)O—.

As used herein, the term “oxycarbonylthio” refers to a divalent group offormula —O(CO)S—.

As used herein, the term “perfluoroalkyl” refers to an alkyl group inwhich all of the hydrogen atoms are replaced with fluorine atoms.Perfluoroalkyl groups are a subset of fluoroalkyl groups.

As used herein, the term “phosphato” refers to a monovalent group offormula —OPO₃H₂.

As used herein, the term “phosphono” refers to a monovalent group offormula —PO₃H₂.

As used herein, the term “phosphoramido” refers to a monovalent group offormula —NHPO₃H₂.

As used herein, the term “primary aromatic amino” refers to a monovalentgroup of formula —ArNH₂ where Ar is an aryl group.

As used herein, the term “secondary aromatic amino” refers to amonovalent group of formula —ArNR^(h)H where Ar is an aryl group andR^(h) is an alkyl or aryl.

As used herein, the term “tertiary amino” refers to a group of formula—NR₂ where R is an alkyl.

As used herein, the term “thio” refers to a divalent group of formula—S—.

As used herein, the term “thiocarbonylimino” refers to a divalent groupof formula —S(CO)NR⁴— where R⁴ is hydrogen, alkyl, or aryl.

As used herein, the term “attachment group” refers to the group formedby reaction of a substrate-reactive group in a compound according toFormula I with a complementary functional group on the surface of asubstrate.

As used herein, the term “complementary functional group” refers to agroup capable of reacting with a recited group to form an ionic bond,covalent bond, or combinations thereof. For example, the complementaryfunctional group can be a group on a substrate capable of reacting withgroup X¹ in Formula I.

As used herein, the term “connector group” refers to a group linking asubstrate to an immobilized amine-containing material. The attachmentgroup is part of the connector group.

As used herein, the term “room temperature” refers to a temperature ofabout 20° C. to about 25° C. or about 22° C. to about 25° C.

As used herein, the term “substrate” refers to a solid phase support towhich the tethering compounds of the invention can be attached. Thesubstrates can have any useful form including, but not limited to, thinfilms, sheets, membranes, filters, nonwoven or woven fibers, hollow orsolid beads, bottles, plates, tubes, rods, pipes, or wafers. Thesubstrates can be porous or non-porous, rigid or flexible, transparentor opaque, clear or colored, or reflective or non-reflective. Suitablesubstrate materials include, for example, polymeric materials, glasses,ceramics, metals, metal oxides, hydrated metal oxides, or combinationsthereof.

As used herein, the term “tethering compound” refers to a compound thathas two reactive groups. One of the reactive groups (i.e., thesubstrate-reactive functional group) can react with a complementaryfunctional group on the surface of a substrate to form a tetheringgroup. The other reactive group (i.e., the N-sulfonylaminocarbonylgroup) can react with an amine-containing material. Reaction of bothreactive groups of the tethering compound results in the formation of aconnector group between the substrate and the amine-containing material(i.e., the amine-containing material can be immobilized on thesubstrate).

As used herein, the term “tethering group” refers to a group attached toa substrate that results from the reaction of a tethering compound witha complementary functional group on the surface of the substrate with atethering compound. The tethering group includes a group that can reactwith an amine-containing material. The tethering group includes aN-sulfonylaminocarbonyl group. The attachment group is part of thetethering group.

As used herein, a curve connecting two groups in a formula indicatesthat the two groups together form part of a cyclic structure.

Compounds

One aspect of the invention provides tethering compounds. The compoundsinclude both a substrate-reactive group and a N-sulfonylaminocarbonylgroup. The substrate-reactive group can be used for attachment to asubstrate and the N-sulfonylaminocarbonyl group can be reacted with anamine-containing material to form a carbonylimino-containing connectorgroup resulting in the immobilization of the amine-containing materialto a substrate. That is, the compounds can be reacted to provide aconnector group between a substrate and an amine-containing material.

The compounds are of Formula I:

wherein

X¹ is a substrate-reactive functional group selected from a carboxy,halocarbonyl, halocarbonyloxy, cyano, hydroxy, mercapto, isocyanato,halosilyl, alkoxysilyl, acyloxysilyl, azido, aziridinyl, haloalkyl,tertiary amino, primary aromatic amino, secondary aromatic amino,disulfide, alkyl disulfide, benzotriazolyl, phosphono, phosphoroamido,phosphato, or ethylenically unsaturated group;

Y¹ is a single bond or a divalent group selected from an alkylene,heteroalkylene, arylene, carbonyl, carbonyloxy, carbonylimino, oxy,thio, —NR^(d)— where R^(d) is hydrogen or alkyl, or combinationsthereof;

Z¹ is an alkyl, aryl, or —(CO)R^(a) wherein R^(a) together with R¹ andgroups to which they are attached form a four to eight memberedheterocyclic or heterobicyclic group having a nitrogen heteroatom and asulfur heteroatom, wherein said heterocyclic or heterobicyclic group canbe fused to an optional aromatic group, optional saturated orunsaturated cyclic group, or optional saturated or unsaturated bicyclicgroup;

R¹ is an alkyl, fluoroalkyl, chloroalkyl, aryl, —NR^(b)R^(c) whereinR^(b) and R^(c) are each an alkyl group or taken together with thenitrogen atom to which they are attached form a four to eight memberedcyclic group, or R¹ together with R^(a) and the groups to which they areattached form the four to eight membered heterocyclic or heterobicyclicgroup that can be fused to the optional aromatic group, optionalsaturated or unsaturated cyclic group, or optional saturated orunsaturated bicyclic group; and

r is equal to 1 when X¹ is a monovalent group or equal to 2 when X¹ is adivalent group. The compounds of Formula I can be unsubstituted orsubstituted with a halo, alkyl, alkoxy, or combinations thereof.

The functional group X¹ typically does not react with a group—(CO)NZ¹SO₂R¹ in Formula I and can be used, for example, to provideattachment to a substrate by reacting with a complementary functionalgroup on the surface of the substrate. That is, X¹ in compounds ofFormula I can react with a complementary functional group to form asubstrate-attached tethering group. X¹ can be monovalent or divalent.When X¹ is divalent, r in Formula I is equal to 2 and the compound hasthe following structure:

The compound can be symmetrical about X¹. A disulfide is an exemplarydivalent X¹ group. When X¹ is monovalent, r in Formula I is equal to 1and the compound has the following structure:

Suitable monovalent X¹ groups include a carboxy, halocarbonyl,halocarbonyloxy, cyano, hydroxy, mercapto, isocyanato, halosilyl,alkoxysilyl, acyloxysilyl, azido, aziridinyl, haloalkyl, tertiary amino,primary aromatic amino, secondary aromatic amino, disulfide, alkyldisulfide, benzotriazolyl, phosphono, phosphoroamido, phosphato, orethylenically unsaturated group.

The X¹ groups typically can react with a complementary functional groupon the surface of a substrate to form an ionic bond, covalent bond, orcombination thereof. Suitable X¹ groups for attachment to the surface ofa polymeric substrate include a carboxy, halocarbonyl, halocarbonyloxy,cyano, hydroxy, mercapto, isocyanato, halosilyl, alkoxysilyl,acyloxysilyl, azido, aziridinyl, haloalkyl, tertiary amino, primaryaromatic amino, secondary aromatic amino, or ethylenically unsaturatedgroup. Suitable X¹ groups for attachment to the surface of agold-containing substrate include mercapto, disulfide, or alkyldisulfide. Suitable X¹ groups for attachment to the surface of othermetal-containing substrates include benzotriazolyl, phosphono,phosphoroamido, or phosphato groups. Suitable X¹ groups for attachmentto glass or ceramic-containing substrates as well as to metaloxide-containing or hydrated metal oxide-containing substrates includehalosilyl, alkoxysilyl, or acyloxysilyl groups.

The group Y¹ in Formula I can be a single bond or a divalent groupselected from an alkylene, heteroalkylene, arylene, carbonyl,carbonyloxy, carbonylimino, oxy, thio, —NR^(d)— where R^(d) is hydrogenor alkyl, or combinations thereof. Suitable heteroalkylenes usuallycontain 1 to 400 carbon atoms and up to 30 heteroatoms selected from N,O, S, or combinations thereof. Suitable alkylenes usually contain 1 to200 carbon atoms. The heteroalkylene and alkylene groups can be linear,branched, cyclic, or combinations thereof.

The group Y¹, for example, can include an alkylene group as in thefollowing formula:

where n is an integer of 1 to 100. Exemplary compounds include thosewhere n is an integer no greater than 80, no greater than 60, no greaterthan 40, no greater than 20, or no greater than 10. The groups X¹, r,Z¹, and R¹ are the same as previously defined for Formula I. Thecompounds can be unsubstituted or substituted with a halo, alkyl,alkoxy, or combinations thereof.

In some compounds according to Formula I, Y¹ includes a first alkylenegroup that is linked to a second alkylene or a first heteroalkylenegroup with a group selected from a carbonyl, carbonyloxy, carbonylimino,oxy, thio, or —NR^(d)— where R^(d) is hydrogen or alkyl. Additionalalkylene or heteroalkylene groups can be linked to the second alkyleneor to the first heteroalkylene group with a group selected from acarbonyl, carbonyloxy, carbonylimino, oxy, thio, or —NR^(d)— where R^(d)is hydrogen or alkyl. In other compounds according to Formula I, Y¹includes a first heteroalkylene group that is linked to a secondheteroalkylene or to a first alkylene group with a group selected from acarbonyl, carbonyloxy, carbonylimino oxy, thio, or —NR^(d)— where R^(d)is hydrogen or alkyl. Additional alkylene or heteroalkylene groups canbe linked to the second heteroalkylene or to the first alkylene groupwith a group selected from a carbonyl, carbonyloxy, carbonylimino group,oxy, thio, —NR^(d)— where R^(d) is hydrogen or alkyl.

For example, compounds can have the following formula:

where D is oxygen, sulfur, or NH; m is an integer of 1 to 200; t is aninteger of 0 to 12; and k is an integer of 2 to 4. Exemplary compoundsinclude those where m is an integer no greater than 150, no greater than100, no greater than 80, no greater than 60, no greater than 40, nogreater than 20, or no greater than 10; t is an integer no greater than10, no greater than 8, no greater than 6, no greater than 4, no greaterthan 2, or equal to 0; and k is no greater than 3, no greater than 2, orequal to 1. In some compounds, the heteroatom D is oxygen and k is equalto 2. The groups X¹, r, Z¹, and R¹ are the same as previously definedfor Formula I. The compounds can be unsubstituted or substituted with ahalo, alkyl, alkoxy, or combinations thereof.

In other examples, the group Y¹ can include a combination of alkyleneand heteroalkylene groups that are separated by a carbonyl, carbonyloxy,carbonylimino, oxy, thio, —NR^(d)— where R^(d) is hydrogen or alkyl, orcombinations thereof:

where D is oxygen, sulfur, or NH; n is an integer of 1 to 100; m is aninteger of 1 to 200; t is an integer of 0 to 12; k is an integer of 2 to4; and L is oxygen or NR^(d) where R^(d) is hydrogen or alkyl. Thepositions of the alkylene groups and the heteroalkylene groups can bereversed. Exemplary compounds include those where n is an integer nogreater than 80, no greater than 60, no greater than 40, no greater than20, or no greater than 10; m is an integer no greater than 150, nogreater than 100, no greater than 80, no greater than 60, no greaterthan 40, no greater than 20, or no greater than 10; t is an integer nogreater than 10, no greater than 8, no greater than 6, no greater than4, no greater than 2, or equal to 0; and k is an integer no greater than3, no greater than 2, or equal to 1. In some compounds, the D group isoxygen and k is equal to 2. The groups X¹, r, Z¹, and R¹ are the same aspreviously defined for Formula I. The compounds can be unsubstituted orsubstituted with a halo, alkyl, alkoxy, or combinations thereof.

In other examples, the compound can have one the following formulas:

where D is oxygen, sulfur, or NH; n is an integer of 1 to 100; m is aninteger of 1 to 200; p is an integer of 1 to 10; t is an integer of 0 to12; k is an integer of 2 to 4; and L is oxygen or NR^(d) where R^(d) ishydrogen or alkyl. Exemplary compounds include those in which n is aninteger no greater than 80, no greater than 60, no greater than 40, nogreater than 20, or no greater than 10; m is an integer no greater than150, no greater than 100, no greater than 80, no greater than 60, nogreater than 40, no greater than 20, or no greater than 10; p is aninteger no greater than 8, no greater than 6, no greater than 4, or nogreater than 2; t is an integer no greater than 10, no greater than 8,no greater than 6, no greater than 4, no greater than 2, or equal to 0;and k is an integer no greater than 3, no greater than 2, or equal to 1.In some compounds, the heteroatom D is oxygen and k is equal to 2. Thegroups X¹, r, Z¹, and R¹ are the same as previously defined for FormulaI. The compounds can be unsubstituted or substituted with a halo, alkyl,alkoxy, or combinations thereof

In some embodiments, Y¹ can include an arylene group in addition to oneor more alkylene groups and one or more heteroalkylene groups. Thearylene can be bonded directly to the N-sulfonylaminocarbonyl group. Thearylene group can include up to 30 carbon atoms, up to 24 carbon atoms,up to 18 carbon atoms, up to 12 carbon atoms, or 6 carbon atoms. In someembodiments, the compounds can have one of the following formulas:

where D is oxygen, sulfur, or NH; n is an integer of 1 to 100; m is aninteger of 1 to 200; p is an integer of 1 to 10; t is an integer of 0 to12; k is an integer of 2 to 4; Ar¹ is an arylene group; and L is oxygenor NR^(d) where R^(d) is hydrogen or alkyl. Exemplary compounds includethose where n is an integer no greater than 80, no greater than 60, nogreater than 40, no greater than 20, or no greater than 10; m is aninteger no greater than 100, no greater than 80, no greater than 60, nogreater than 40, no greater than 20, or no greater than 10; p is aninteger no greater than 8, no greater than 6, no greater than 4, or nogreater than 2; t is an integer no greater than 10, no greater than 8,no greater than 6, no greater than 4, no greater than 2, or equal to 0;and k is an integer no greater than 3, no greater than 2, or equal to 1.In some compounds of the above formula, D is oxygen, k is equal to 2,and Ar¹ is phenylene as in one of the following structures:

The groups X¹, r, Z¹, and R¹ are the same as previously defined forFormula I. The compounds can be unsubstituted or substituted with ahalo, alkyl, alkoxy, or combinations thereof.

The group Y¹ can be an arylene group as in the following structure:

where Ar¹ is an arylene group. In some compounds, Ar¹ is phenylene. Thegroups X¹, r, Z¹, and R¹ are the same as previously defined for FormulaI. The compounds can be unsubstituted or substituted with a halo, alkyl,alkoxy, or combinations thereof.

Y¹ can be a single bond. For example, in the following formula, Y is asingle bond when X¹ is a primary or secondary aromatic amino group.

In this formula, R⁴ is hydrogen, aryl, alkyl. The groups Z¹ and R¹ arethe same as previously defined for Formula I. The compounds can beunsubstituted or substituted with a halo, alkyl, alkoxy, or combinationsthereof.

The group Z¹ in some embodiments of Formula I can be alkyl or aryl. Forexample, Z¹ can be a C₁₋₃₀ alkyl, a C₁₋₁₀ alkyl, or a C₁₋₆ alkyl. Inother examples, Z¹ can be a C₆₋₃₀ aryl, a C₆₋₂₄ aryl, a C₆₋₁₈ aryl, or aC₆₋₁₂ aryl. In other embodiments of Formula I, Z¹ can be a —(CO)R^(a)group that together with R¹ and the groups to which they are attachedform a heterocyclic or heterobicyclic group that can be fused to anoptional aromatic group, optional saturated or unsaturated cyclic group,or optional saturated or unsaturated bicyclic group. The heterocyclic orheterobicyclic group includes a nitrogen and sulfur heteroatom. Anexemplary heterocyclic group fused to an aromatic group is shown in thefollowing formula:

where X¹ is monovalent or

where X¹ is divalent.

As alternatives to combining with Z¹ to form a heterocyclic orheterobicyclic structure, R¹ can be an alkyl, fluoroalkyl, chloroalkyl,aryl, or —NR^(b)R^(c) group where Rb and R_(c) are each an alkyl ortaken together with the nitrogen atom to which they are attached form afour to eight membered heterocyclic group. In some embodiments, R¹ canbe a C₁₋₃₀ alkyl, a C₁₋₁₀ alkyl, or a C₁₋₆ alkyl. In other embodiments,R¹ can be a C₁₋₃₀ fluoroalkyl, a C₁₋₁₀ fluoroalkyl, or a C₁₄perfluoroalkyl group. In still other embodiments, R¹ can be a C₆₋₃₀aryl, a C₆₋₁₈ aryl, or a C₆₋₁₂ aryl. For example R¹ can be a phenylgroup.

Exemplary compounds according to Formula I include, but are not limitedto, the following:

Any of these compounds or any other compound that is within the scope ofFormula I can be unsubstituted or substituted with a halo, alkyl,alkoxy, or combinations thereof.Method of Preparing Compounds

The compounds of Formulas I may be prepared, for example, by reaction ofa first compound having a nitrogen-containing group with a secondcompound that includes a halocarbonyl group. More specifically, thenitrogen-containing group of the first compound includes a nitrogen atomdirectly bonded to a sulfonyl group as well as to at least one hydrogenatom. The first compound can further include a substrate-reactive groupX¹ or a group that can be converted to a substrate-reactive group X¹.The substrate-reactive groups do not react, or react slowly, with thehalocarbonyl group of the second compound such that thenitrogen-containing group of the first compound reacts preferentiallywith the halocarbonyl group of the second compound. A suitable reactionscheme is shown in Reaction Scheme A.

The —(CO)Q group can be a halocarbonyl group. The groups X¹, Y¹, Z¹, andR¹ can be the same as previously defined for Formula I. Where Z¹ inFormula I is the —(CO)R^(a) and R^(a) combines with R¹ to form a ringstructure, the compounds can be prepared using reaction Scheme A′.

where R^(a), R¹, X¹, and Y¹ are as previously defined for Formula I. The—(CO)Q group can be a halocarbonyl group.Articles

Another aspect of the invention provides articles that include atethering group attached to a substrate (i.e., a substrate-attachedtethering group). The substrate-attached tethering group is the reactionproduct of a complementary functional group G on a surface of asubstrate with the group X¹ in compounds of Formula I. Thesubstrate-attached tethering group has a N-sulfonylaminocarbonyl groupthat can react with an amine-containing material to form acarbonylimino-containing connector group between a substrate and anamine-containing material.

The substrate is a solid phase material to which the tethering groupscan be attached. The substrate is not soluble in a solution used toattach a compound of Formula I to the surface of the substrate.Typically, a tethering group is attached only to an outer portion of thesubstrate and a bulk portion of the substrate is not modified during theprocess of attaching tethering group to the substrate. If the substratehas groups G distributed throughout the substrate, only those groups inthe outer portion (e.g., on or near the surface) are usually capable ofreacting with group X¹ of the compounds according to Formula I.

The substrates can have any useful form including, but not limited to,thin films, sheets, membranes, filters, nonwoven or woven fibers, hollowor solid beads, bottles, plates, tubes, rods, pipes, or wafers. Thesubstrates can be porous or non-porous, rigid or flexible, transparentor opaque, clear or colored, and reflective or non-reflective. Suitablesubstrate materials include, for example, polymeric materials, glasses,ceramics, metals, metal oxides, hydrated metal oxides, or combinationsthereof.

The substrates can have a single layer or multiple layers of material.For example, the substrate can have one or more second layers thatprovide support for a first layer that includes a complementaryfunctional group capable of reacting with the X¹ group in compound ofFormula I. The first layer is the outer layer of the substrate. In someembodiments, a surface of a second layer is chemically modified orcoated with another material to provide a first layer that includes acomplementary functional group capable of reacting with the X¹ group.

Suitable polymeric substrate materials include, but are not limited to,polyolefins, polystyrenes, polyacrylates, polymethacrylates,polyacrylonitriles, poly(vinylacetates), polyvinyl alcohols, polyvinylchlorides, polyoxymethylenes, polycarbonates, polyamides, polyimides,polyurethanes, phenolics, polyamines, amino-epoxy resins, polyesters,silicones, cellulose based polymers, polysaccharides, or combinationsthereof. In some embodiments, the polymeric material is a copolymerprepared using a comonomer having a complementary functional groupcapable of reacting with a group X¹ in compounds according to Formula I.For example, the comonomer can contain a carboxy, mercapto, hydroxy,amino, or alkoxysilyl group.

Suitable glass and ceramic substrate materials can include, for example,sodium, silicon, aluminum, lead, boron, phosphorous, zirconium,magnesium, calcium, arsenic, gallium, titanium, copper, or combinationsthereof. Glasses typically include various types of silicate containingmaterials.

In some embodiments, the substrate includes a layer of diamond-likeglass as disclosed in International Patent Application WO 01/66820 A1,the disclosure of which is incorporated herein by reference in itsentirety. The diamond-like glass is an amorphous material that includescarbon, silicon, and one or more elements selected from hydrogen,oxygen, fluorine, sulfur, titanium, or copper. Some diamond-like glassmaterials are formed from a tetramethysilane precursor using a plasmaprocess. A hydrophobic material can be produced that is further treatedin an oxygen plasma to control the silanol concentration on the surface.

Diamond-like glass can be in the form of a thin film or in the form of acoating on another layer or material in the substrate. In someapplications, the diamond-like glass can be in the form of a thin filmhaving at least 30 weight percent carbon, at least 25 weight percentsilicon, and up to 45 weight percent oxygen. Such films can be flexibleand transparent. In some embodiments, the diamond-like glass is theouter layer of a multilayer substrate. In a specific example, the secondlayer (e.g., support layer) of the substrate is a polymeric material andthe first layer is a thin film of diamond-like glass. The tetheringgroup is attached to the surface of the diamond-like glass.

In some multilayer substrates, the diamond like glass is deposited on alayer of diamond-like carbon. For example, the second layer (e.g.,support layer) is a polymeric film having a layer of diamond-like carbondeposited on a surface. A layer of diamond-like glass is deposited overthe diamond-like carbon layer. The diamond-like carbon can, in someembodiments, function as a tie layer or primer layer between a polymericlayer and a layer of diamond-like glass in a multilayer substrate. Forexample, the multilayer substrate can include a polyimide or polyesterlayer, a layer of diamond-like carbon deposited on the polyimide orpolyester, and a layer of diamond-like glass deposited on thediamond-like carbon. In another example, the multilayer substrateincludes a stack of the layers arranged in the following order:diamond-like glass, diamond-like carbon, polyimide or polyester,diamond-like carbon, and diamond-like glass.

Diamond-like carbon films can be prepared, for example, from acetylenein a plasma reactor. Other methods of preparing such films are describedU.S. Pat. Nos. 5,888,594 and 5,948,166 as well as in the article M.David et al., AlChE Journal, 37 (3), 367-376 (March 1991), thedisclosures of which are incorporated herein by reference.

Suitable metals, metal oxides, or hydrated metal oxides for substratescan include, for example, gold, silver, platinum, palladium, aluminum,copper, chromium, iron, cobalt, nickel, zinc, and the like. Themetal-containing material can be alloys such as stainless steel, indiumtin oxide, and the like. In some embodiments, a metal-containingmaterial is the top layer of a multilayer substrate. For example, thesubstrate can have a polymeric second layer and a metal containing firstlayer. In one example, the second layer is a polymeric film and thefirst layer is a thin film of gold. In other examples, a multilayersubstrate includes a polymeric film coated with a titanium-containinglayer and then coated with a gold-containing layer. That is, thetitanium layer can function as a tie layer or a primer layer foradhering the layer of gold to the polymeric film.

In other examples of a multilayer substrate, a silicon support layer iscovered with a layer of chromium and then with a layer of gold. Thechromium layer can improve the adhesion of the gold layer to the siliconlayer.

The surface of the substrate typically includes a group capable ofreacting with a carboxy, halocarbonyl, halocarbonyloxy, cyano, hydroxy,mercapto, isocyanato, halosilyl, alkoxysilyl, acyloxysilyl, azido,aziridinyl, haloalkyl, tertiary amino, primary aromatic amino, secondaryaromatic amino, disulfide, alkyl disulfide, benzotriazolyl, phosphono,phosphoroamido, phosphato, or ethylenically unsaturated group. That is,the substrate includes a group capable of reacting with the group X¹ incompounds of Formula I (i.e., the substrate includes a complementaryfunctional group to the group X¹). Substrates can include a supportmaterial that is treated to have an outer layer that includes acomplementary functional group. The substrate can be prepared from anysolid phase material known to have groups capable of reacting with X¹and is not limited to the following examples of suitable materials.

A carboxy group or a halocarbonyl group can react with a substratehaving a hydroxy group to form a carbonyloxy-containing attachmentgroup. Examples of substrate materials having hydroxy groups include,but are not limited to, polyvinyl alcohol, corona-treated polyethylene,hydroxy substituted esters of polymethacrylates, hydroxy substitutedesters of polyacrylates, and a polyvinyl alcohol coating on a supportmaterial such as glass or polymer film.

A carboxy group or a halocarbonyl group can also react with a substratehaving a mercapto group to form a carbonylthio-containing attachmentgroup. Examples of substrate materials having a mercapto group include,but are not limited to, mercapto substituted esters of polyacrylates,mercapto substituted esters of polymethacrylates, and glass treated witha mercaptoalkylsilane.

Additionally, a carboxy group or a halocarbonyl group can react with aprimary aromatic amino group, a secondary aromatic amino group, or asecondary aliphatic amino group to form a carbonylimino-containingattachment group. Examples of substrate materials having aromaticprimary or secondary amino groups include, but are not limited to,polyamines, amine substituted esters of polymethacrylate, aminesubstituted esters of polyacrylate, polyethylenimines, and glass treatedwith an aminoalkylsilane.

A halocarbonyloxy group can react with a substrate having a hydroxygroup to form an oxycarbonyloxy-containing attachment group. Examples ofsubstrate materials having hydroxy groups include, but are not limitedto, polyvinyl alcohol, corona-treated polyethylene, hydroxy substitutedesters of polymethacrylates, hydroxy substituted esters ofpolyacrylates, and a polyvinyl alcohol coating on a support materialsuch as glass or polymer film.

A halocarbonyloxy group can also react with a substrate having amercapto group to form an oxycarbonylthio-containing attachment group.Examples of substrate materials having a mercapto group include, but arenot limited to, mercapto substituted esters of polymethacrylates,mercapto substituted esters of polyacrylates, and glass treated with amercaptoalkylsilane.

Additionally, a halocarbonyloxy group can react with a substrate havinga primary aromatic amino group, a secondary aromatic amino group, or asecondary aliphatic amino group to form an oxycarbonylimino-containingattachment group. Examples of substrate materials having aromaticprimary or secondary amino groups include, but are not limited to,polyamines, amine substituted esters of polymethacrylate, aminesubstituted esters of polyacrylate, polyethylenimines, and glass treatedwith an aminoalkylsilane.

A cyano group can react with a substrate having an azido group to form atetrazinediyl-containing attachment group. Examples of substrates havingazido groups include, but are not limited to, a coating ofpoly(4-azidomethylstyrene) on a glass or polymeric support. Suitablepolymeric support materials include polyesters, polyimides, and thelike.

A hydroxy group can react with a substrate having isocyanate group toform an oxycarbonylimino-containing attachment group. Suitablesubstrates having isocyanate groups include, but are not limited to, acoating of 2-isocyanatoethylmethacrylate polymer on a support material.Suitable support materials include glass and polymeric materials such aspolyesters, polyimides, and the like.

A hydroxy group can also react with a substrate having a carboxy,carbonyloxycarbonyl, or halocarbonyl to form a carbonyloxy-containingattachment group. Suitable substrates include, but are not limited to, acoating of acrylic acid polymer or copolymer on a support material or acoating of a methacrylic acid polymer or copolymer on a supportmaterial. Suitable support materials include glass and polymericmaterials such as polyesters, polyimides, and the like. Other suitablesubstrates include copolymers of polyethylene with polyacrylic acid,polymethacrylic acid, or combinations thereof.

A mercapto group can react with a substrate having isocyanate groups.The reaction between a mercapto group and an isocyanate group forms athiocarbonylimino-containing attachment group. Suitable substrateshaving isocyanate groups include, but are not limited to, a coating of2-isocyanatoethylmethacrylate copolymer on a support material. Suitablesupport materials include glass and polymeric materials such aspolyesters, polyimides, and the like.

A mercapto group can also react with a substrate having a halocarbonylgroup to form a carbonylthio-containing attachment group. Substrateshaving halocarbonyl groups include, for example, chlorocarbonylsubstituted polyethylene.

A mercapto group can also react with a substrate having ahalocarbonyloxy group to form an oxycarbonlythio-containing attachmentgroup. Substrates having halocarbonyl groups include chloroformyl estersof polyvinyl alcohol.

Additionally, a mercapto group can react with a substrate having anethylenically unsaturated group to form a thioether-containingattachment group. Suitable substrates having an ethylenicallyunsaturated group include, but are not limited to, polymers andcopolymers derived from butadiene.

An isocyanate group can react with a substrate having a hydroxy group toform a oxycarbonylimino-containing attachment group. Examples ofsubstrate materials having hydroxy groups include, but are not limitedto, polyvinyl alcohol, corona-treated polyethylene, hydroxy substitutedesters of polymethacrylates or polyacrylates, and a polyvinyl alcoholcoating on glass or polymer film.

An isocyanate group can also react with a mercapto group to form athiocarbonylimino-containing attachment group. Examples of substratematerials having a mercapto group include, but are not limited to,mercapto substituted esters of polymethacrylates or polyacrylates andglass treated with a mercaptoalkylsilane.

Additionally, an isocyanate group can react with a primary aromaticamino group, a secondary aromatic amino group, or a secondary aliphaticamino group to form a urea-containing attachment group. Suitablesubstrates having a primary or secondary aromatic amino group include,but are not limited to, polyamines, polyethylenimines, and coatings ofan aminoalkylsilane on a support material such as glass or on apolymeric material such as a polyester or polyimide.

An isocyanate group can also react with a carboxy to form an O-acylcarbamoyl-containing attachment group. Suitable substrates having acarboxylic acid group include, but are not limited to, a coating of anacrylic acid polymer or copolymer or a coating of a methacrylic acidpolymer or copolymer on a glass or polymeric support. Copolymersinclude, but are not limited to, copolymers that contain polyethyleneand polyacrylic acid or polymethacrylic acid. Suitable polymeric supportmaterials include polyesters, polyimides, and the like.

A halosilyl group, an alkoxysilyl group, or an acyloxysilyl group canreact with a substrate having a silanol group to form adisiloxane-containing attachment group. Suitable substrates includethose prepared from various glasses, ceramic materials, or polymericmaterial. These groups can also react with various materials havingmetal hydroxide groups on the surface to form a silane-containinglinkage. Suitable metals include, but are not limited to, silver,aluminum, copper, chromium, iron, cobalt, nickel, zinc, and the like. Insome embodiments, the metal is stainless steel or another alloy.Polymeric material can be prepared to have silanol groups. For example,commercially available monomers with silanol groups include3-(trimethoxysilyl)propyl methacrylate and 3-aminoproplytrimethoxysilaneavailable from Aldrich Chemical Co., Milwaukee, Wis.

An azido group can react, for example, with a substrate havingcarbon-carbon triple bond to form triazolediyl-containing attachmentgroups. An azido group can also react with a substrate having nitrilegroups to form a tetrazenediyl-containing attachment group. Substrateshaving nitrile groups include, but are not limited to, coatings ofpolyacrylonitrile on a support material such as glass or a polymericmaterial. Suitable polymeric support material includes polyesters andpolyimides, for example. Other suitable substrates having nitrile groupsinclude acrylonitrile polymers or copolymers and 2-cyanoacrylatepolymers or copolymers.

An azido group can also react with a strained olefinic group to form atriazolediyl-containing attachment group. Suitable substrates have astrained olefinic group include coatings that have pendant norbornenylfunctional groups. Suitable support materials include, but are notlimited to, glass and polymeric materials such as polyesters andpolyimides.

An aziridinyl group can react with a mercapto group to form aaminoalkylthioether-containing attachment group. Examples of substratematerials having a mercapto group include, but are not limited to,mercapto substituted esters of polymethacrylates or polyacrylates andglass treated with a mercaptoalkylsilane.

Additionally, an aziridinyl group can react with a carboxy group to forma aminoalkyloxycarbonyl-containing attachment group. Suitable substrateshaving a carboxy include, but are not limited to, a coating of a acrylicacid polymer or copolymer, or a coating of a methacrylic acid polymer orcopolymer on a glass or polymeric support. Copolymers include, but arenot limited to, copolymers that contain polyethylene and polyacrylicacid or polymethacrylic acid. Suitable polymeric support materialsinclude polyesters, polyimides, and the like.

A haloalkyl group can react, for example, with a substrate having atertiary amino group to form a quaternary ammonium-containing attachmentgroup. Suitable substrates having a tertiary amino group include, butare not limited to, polydimethylaminostyrene orpolydimethylaminoethylmethacrylate.

Likewise, a tertiary amino group can react, for example, with asubstrate having a haloalkyl group to form a quaternaryammonium-containing attachment group. Suitable substrates having ahaloalkyl group include, for example, coatings of a haloalkylsilane on asupport material. Support materials can include, but are not limited to,glass and polymeric materials such as polyesters and polyimides.

A primary aromatic amino or a secondary aromatic amino group can react,for example, with a substrate having an isocyanate group to form aoxycarbonylimino-containing attachment group. Suitable substrates havingisocyanate groups include, but are not limited to, a coating of a2-isocyanatoethylmethacrylate polymer or copolymer on a glass orpolymeric support. Suitable polymeric supports include polyesters,polyimides, and the like.

A primary aromatic amino or a secondary aromatic amino group can alsoreact with a substrate containing a carboxy or halocarbonyl group toform a carbonylimino-containing attachment group. Suitable substratesinclude, but are not limited to, acrylic or methacrylic acid polymericcoatings on a support material. The support material can be, forexample, glass or a polymeric material such as polyesters or polyimides.Other suitable substrates include copolymers of polyethylene andpolymethacrylic acid or polyacrylic acid.

A disulfide or an alkyl disulfide group can react, for example, with ametal surface to form a metal sulfide-containing attachment group.Suitable metals include, but are not limited to gold, platinum,palladium, nickel, copper, and chromium. The substrate can also be analloy such an indium tin oxide or a dielectric material.

A benzotriazolyl can react, for example, with a substrate having a metalor metal oxide surface. Suitable metals or metal oxides include, forexample, silver, aluminum, copper, chromium, iron, cobalt, nickel, zinc,and the like. The metals or metal oxides can include alloys such asstainless steel, indium tin oxide, and the like.

A phosphono, phosphoroamido, or phosphate can react, for example, with asubstrate having a metal or metal oxide surface. Suitable metals ormetal oxides include, for example, silver, aluminum, copper, chromium,iron, cobalt, nickel, zinc, and the like. The metals or metal oxides caninclude alloys such as stainless steel, indium tin oxide, and the like.

An ethylenically unsaturated group can react, for example, with asubstrate having an alkyl group substituted with a mercapto group. Thereaction forms a heteroalkylene-containing attachment group. Suitablesubstrates include, for example, mercapto-substituted alkyl esters ofpolyacrylates or polymethacrylates.

An ethylenically unsaturated group can also react with a substratehaving a silicon surface, such as a silicon substrate formed using achemical vapor deposition process. Such silicon surfaces can contain—SiH groups that can react with the ethylenically unsaturated group inthe presence of a platinum catalyst to form an attachment group with Sibonded to an alkylene group.

Additionally, an ethylenically unsaturated group can react with asubstrate having a carbon-carbon double bond to form analkylene-containing attachment group. Such substrates include, forexample, polymers derived from butadiene.

The compounds of Formula I can undergo a self-assembly process whencontacted with a substrate. As used herein, the term “self-assembly”refers to process in which a material can spontaneously form a monolayerof substrate-attached tethering groups when contacted with a substrate.For example, compounds having a disulfide or alkyl disulfide group forX¹ can undergo a self-assembly process when exposed to a gold substrate.As another example, compounds having a halosilyl group for X¹ canundergo a self-assembly process when exposed to a diamond-like glass orglass substrate.

In one embodiment, Formula II can represent the articles of theinvention:

Formula II represents a tethering group attached to a substrate. Thetethering group is derived from a compound according to Formula I. Thegroup U¹ is the attachment group formed by reaction of X¹ in a compoundaccording to Formula I with a complementary functional group on thesurface of a substrate. The groups Y¹ and R¹ are the same as previouslydefined for Formula I. That is, the article includes:

a substrate; and

a substrate-attached tethering group that includes a reaction product ofa complementary functional group G on a surface of the substrate with acompound of Formula I

wherein

X¹ is a substrate-reactive functional group selected from a carboxy,halocarbonyl, halocarbonyloxy, cyano, hydroxy, mercapto, isocyanato,halosilyl, alkoxysilyl, acyloxysilyl, azido, aziridinyl, haloalkyl,tertiary amino, primary aromatic amino, secondary aromatic amino,disulfide, alkyl disulfide, benzotriazolyl, phosphono, phosphoroamido,or phosphato;

Y¹ is a single bond or a divalent group selected from an alkylene,heteroalkylene, arylene, carbonyl, carbonyloxy, carbonylimino, oxy,thio, —NR^(d)— where R^(d) is hydrogen or alkyl, or combinationsthereof;

Z¹ is an alkyl, aryl, or —(CO)R^(a) wherein R^(a) together with R¹ andgroups to which they are attached form a four to eight memberedheterocyclic or heterobicyclic group having a nitrogen heteroatom and asulfur heteroatom, wherein said heterocyclic or heterobicyclic group canbe fused to an optional aromatic group, optional saturated orunsaturated cyclic group, or optional saturated or unsaturated bicyclicgroup;

R¹ is an alkyl, fluoroalkyl, chloroalkyl, aryl, NR^(b)R^(c) whereinR^(b) and R^(c) are each an alkyl group or taken together with thenitrogen atom to which they are attached form a four to eight memberedcyclic group having a nitrogen heteroatom, or R¹ together with R^(a) andthe groups to which they are attached form the four to eight memberedheterocyclic or heterobicyclic group that can be fused to the optionalaromatic group, optional saturated or unsaturated cyclic group, oroptional saturated or unsaturated bicyclic group;

r is equal to 1 when X¹ is a monovalent group or equal to 2 when X¹ is adivalent group; and

G is a group capable of reacting with X¹ to form an ionic bond, covalentbond, or combinations thereof. The tethering group can be unsubstitutedor substituted with a halo, alkyl, alkoxy, or combinations thereof.

Formula II shows only one tethering group attached to the substrate;however, more than one tethering group can be attached to the substrateif there are more than one reactive group G on the substrate. Further,the substrate can have excess G groups on the surface of the substratethat have not reacted with a tethering compound.

Groups on a substrate (i.e., groups G) capable of reacting with X¹groups in compounds according to Formula I include, but are not limitedto, hydroxy, mercapto, primary aromatic amino group, secondary aromaticamino group, secondary aliphatic amino group, azido, carboxy,carbonyloxycarbonyl, isocyanate, halocarbonyl, halocarbonyloxy, silanol,and nitrile.

The attachment of tethering groups to the surface of a substrate (i.e.,formation of the substrate-attached tethering groups of Formulas II) canbe detected using techniques such as, for example, contact anglemeasurements of a liquid on the substrate before and after attachment ofa tethering group derived from Formula I (e.g., the contact angle canchange upon attachment of a tethering group to the surface of asubstrate), ellipsometry (e.g., the thickness of the attached layer canbe measured), time-of-flight mass spectroscopy (e.g., the surfaceconcentration can change upon attachment of a tethering group to asubstrate), and Fourier Transform Infrared Spectroscopy (e.g., thereflectance and absorbance can change upon attachment of a tetheringgroup to a substrate).

In other embodiments of articles of the invention, theN-sulfonylaminocarbonyl group in the tethering group has reacted with anamine-containing material. A carbonylimino-containing connector group isformed resulting in the immobilization of an amine-containing materialto the substrate. The amine-containing material can react with aN-sulfonylaminocarbonyl group of the substrate-attached tethering groupof Formula II. In some embodiments, the amine-containing materials arebiomolecules such as, for example, amino acid, peptide, DNA, RNA,protein, enzyme, organelle, immunoglobin, or fragments thereof. In otherembodiments, the amine-containing material is a non-biological aminesuch as an amine-containing analyte.

The amine-containing material (H₂N-T) can react with thesubstrate-attached tethering group of Formula II by a nucleophilicsubstitution reaction to produce a substrate immobilizedamine-containing material of Formula III:

where U¹ is the attachment group formed by reacting X¹ of a compoundaccording to Formula I with a complementary functional group on thesurface of the substrate; T is the remainder of the amine-containingmaterial; and Y¹ and R¹ are the same as previously defined for FormulasI and II. H₂N-T is any suitable primary amine-containing material. Insome embodiments, H₂N-T is a biomolecule.

The presence of the immobilized amine can be determined, for example,using mass spectroscopy, contact angle measurement, infraredspectroscopy, and ellipsometry. Additionally, various immunoassays andoptical microscopic techniques can be used if the amine-containingmaterial is a biologically active material.

Other materials can be bound to the amine-containing material. Forexample, a complementary RNA or DNA fragment can hybridize with animmobilized RNA or DNA fragment. In another example, an antigen can bindto an immobilized antibody or an antibody can bind to an immobilizedantigen. In a more specific example, a bacterium such as Staphylococcusaureus can bind to an immobilized biomolecule.

Method of Immobilizing Amine-Containing Material to a Substrate

Another aspect of the invention provides methods for immobilizing anamine-containing material to a substrate. The method involves preparinga substrate-attached tethering group by reacting a complementaryfunctional group on the surface of the substrate with thesubstrate-reactive group X¹ in compounds of Formula I; and reacting aN-sulfonylaminocarbonyl group of the substrate-attached tethering groupwith an amine-containing material to form a carbonylimino-containingconnector group between the substrate and the amine-containing material.

In one embodiment, the method of immobilizing an amine-containingmaterial to a substrate is shown in Reaction Scheme B for a monovalentX¹.

where U¹ is the attachment group formed by reacting X¹ in compound ofFormula I with a complementary functional group G on the surface of thesubstrate; T is the remainder of the amine-containing material, (i.e.,the group T represents all of the amine-containing material exclusive ofthe amine group). The groups Y¹ and R¹ are the same as previouslydefined for Formula I. H₂N-T is any suitable amine-containing material.In some embodiments, H₂N-T is a biomolecule. The method involves:

selecting a compound of Formula I

wherein

X¹ is a substrate-reactive functional group selected from a carboxy,halocarbonyl, halocarbonyloxy, cyano, hydroxy, mercapto, isocyanato,halosilyl, alkoxysilyl, acyloxysilyl, azido, aziridinyl, haloalkyl,tertiary amino, primary aromatic amino, secondary aromatic amino,disulfide, alkyl disulfide, benzotriazolyl, phosphono, phosphoroamido,phosphato, or ethylenically unsaturated group;

Y¹ is a single bond or a divalent group selected from an alkylene,heteroalkylene, arylene, carbonyl, carbonyloxy, carbonylimino, oxy,thio, —NR^(d)— where R^(d) is hydrogen or alkyl, or combinationsthereof;

Z¹ is an alkyl, aryl, or —(CO)R^(a) wherein R^(a) together with R¹ andgroups to which they are attached form a four to eight memberedheterocyclic or heterobicyclic group having a nitrogen heteroatom and asulfur heteroatom, wherein said heterocyclic or heterobicyclic group canbe fused to an optional aromatic group, optional saturated orunsaturated cyclic group, or optional saturated or unsaturated bicyclicgroup;

R¹ is an alkyl, fluoroalkyl, chloroalkyl, aryl, NR^(b)R^(c) whereinR^(b) and R^(c) are each an alkyl group or taken together with thenitrogen atom to which they are attached form a four to eight memberedcyclic group, or R¹ together with R^(a) and the groups to which they areattached form the four to eight membered heterocyclic or heterobicyclicgroup that can be fused to the optional aromatic group, optionalsaturated or unsaturated cyclic group, or optional saturated orunsaturated bicyclic group;

r is equal to 1 when X¹ is a monovalent group or equal to 2 when X¹ is adivalent group; and

said compound of Formula I is unsubstituted or substituted with a halo,alkyl, alkoxy, or combinations thereof;

providing a substrate having a complementary functional group capable ofreacting with X¹;

preparing a substrate-attached tethering group by reacting X¹ with thecomplementary functional group on the substrate resulting in an ionicbond, covalent bond, or combinations thereof, and

reacting a N-sulfonylaminocarbonyl group of the substrate-attachedtethering group with an amine-containing material to form acarbonylimino-containing connector group. That is, the connector groupis a divalent group of formula —U¹—Y¹—(CO)—NH— (the divalent groupbetween the substrate and the group T in Formula III). The attachmentgroup is part of the connector group.

Uses

The compounds of the invention can be used, for example, forimmobilizing amine-containing material. In some embodiments, theamine-containing material is an amine-containing analyte. In otherembodiments, the amine-containing materials are biomolecules such as,for example, amino acids, peptides, DNA, RNA, protein, enzymes,organelles, immunoglobins, or fragments thereof. Immobilized biologicalamine-containing materials can be useful in the medical diagnosis of adisease or of a genetic defect. The immobilized amine-containingmaterials can also be used for biological separations or for detectionof the presence of various biomolecules. Additionally, the immobilizedamine-containing materials can be used in bioreactors or as biocatalyststo prepare other materials. The substrate-attached tethering groups canbe used to detect amine-containing analytes.

Biological amine-containing materials often can remain active afterattachment to the substrate (i.e., the articles according to Formula IIIcan include biologically active amine-containing materials immobilizedto the substrate). For example, an immobilized antibody can bind withantigen or an immobilized antigen can bind to an antibody. Anamine-containing material can bind to a bacterium. In a more specificexample, the immobilized amine-containing material can bind to aStaphylococcus aureus bacterium (e.g., the immobilized amine-containingmaterial can be a biomolecule that has a portion that can specificallybind to the bacterium).

The articles prepared by attaching the compounds of the invention to asubstrate typically have improved hydrolytic stability compared topreviously known articles prepared using a tethering compound that is aderivative of N-hydroxysuccinimide. Because of the hydrolytic stability,the compounds and the substrate-attached tethering groups of theinvention can typically be used in aqueous systems.

When an amine-containing material reacts with a N-sulfonylaminocarbonylgroup, a carbonylimino-containing connector group is formed that resultsin the immobilization of the amine-containing material to the substrate(i.e., substrate immobilized amine-containing materials according toFormulas III). The rate of reaction of amine-containing materials withthe N-sulfonylaminocarbonyl groups of the substrate-attached tetheringgroups is typically faster than the rate of hydrolysis of theN-sulfonylaminocarbonyl group. That is, immobilization ofamine-containing materials occurs at a faster rate than the hydrolysisreactions. The amine-containing materials are not easily displaced onceimmobilization to a substrate has occurred due to the formation of acovalent carbonylimino bond.

EXAMPLES

Unless otherwise noted, all solvents and chemical reagents were or canbe obtained from Aldrich Chemical Co., Milwaukee, Wis.

Gold-coated silicon substrates were obtained from WaferNet, Inc., SanJose, Calif. and were 150 mm prime-grade N-type silicon wafers onto oneside of which metal was deposited by reactive sputtering. The waferswere first treated to deposit, by reactive sputtering, a layer ofchromium and were then treated to deposit, by reactive sputtering, alayer of gold. The thickness of the gold layer was 5000 Angstroms.

N-phenyltrifluoromethanesulfonamide can be obtained from Interchim,Montlucon, France.

Aqueous buffer solutions were obtained from Sigma Aldrich Co.,Milwaukee, Wis. or were prepared by known methods.

HSA and conjugated and unconjugated IgG (i.e., Immunoglobulin G) wereobtained from Jackson ImmunoResearch Laboratories, Inc., West Grove, Pa.

The IR and ¹H NMR spectra of each product formed in the PreparativeExamples and Examples were consistent with the assigned structure.

Glossary

As used herein:

“CHES buffer” refers to an aqueous solution of2-(cyclohexylamino)ethanesulfonic acid;

“DLC” refers to a diamond-like carbon coating prepared as described;

“DLG” refers to a diamond-like glass coating prepared as described;

“DMF” refers to N,N-dimethylformamide;

“ELISA” refers to enzyme-linked immunoabsorbent assay;

“FITC-ALBUMIN” refers to fluorescein-labeled bovine serum albumin, whichwas obtained as a 2 mg/mL solution in bicarbonate buffer fromSigma-Aldrich Corp., St. Louis, Mo.;

“HSA” refers to human serum albumin;

“SA-HRP” refers to streptavidin conjugated with horseradish peroxidase,which was obtained from Jackson ImmunoResearch Laboratories, Inc., WestGrove, Pa.;

“ABTS” refers to 2,2′-azino-di-(3-ethylbenzthiazoline-6-sulfonate),which was obtained in kit form from KPL Inc., Gaithersburg, Md.;

“PBS” refers to phosphate buffered saline which has a pH of about 7.4;

“SDS” refers to sodium dodecyl sulfate; and

“TWEEN 20” refers to polyoxyethylene(20) sorbitan monolaurate.

Methods

Contact Angle Measurements

Advancing and receding contact angles of deionized water were measuredin air at room temperature using a Model 100 goniometer (available fromRame-Hart, Inc., Mountain Lakes, N.J.).

Ellipsometry

Ellipsometric determination of monolayer thickness was carried out usinga Model AutoEL ellipsometer (available from Rudolph Technologies, Inc.,Flanders, N.J.) at a wavelength of 6320 Angstroms and at an angle ofincidence of 70 degrees. For each substrate, ellipsometric constantswere determined by extrapolation of self assembled monolayers of1-mercaptohexadecane, 1-mercaptododecane, and 1-mercaptooctane. Theellipsometric thicknesses of the monolayers were estimated by using athree-layer model and by assuming the refractive index of 1.46 for themonolayer.

Preparative Example 1 Preparation of N-methyltrifluoromethanesulfonamide

A weighed pressure reactor (available from Parr Instrument Co., Moline,Ill.) is charged with dichloromethane (100 mL) and is then cooled usingliquid nitrogen. Methylamine is introduced from a cylinder via astainless steel tube that is connected to a valve on the reactor. Thereactor is periodically weighed, and methylamine is added until 20 ghave been added. Trifluoromethanesulfonylfluoride is then introducedinto the reactor from a cylinder via a stainless steel tube until 97.9 ghave been added. The pressure reactor is then sealed and is placed in amotorized rocker and is rocked and allowed to warm to room temperature.After a period of about 6 hours after the reactor reaches roomtemperature, it is slowly vented to the atmosphere by opening the valve.The product residue is washed with 10 weight percent aqueous HCl and theorganic phase is then dried over MgSO₄. The mixture is filtered and thesolvent is removed from the filtrate using a rotary evaporator to affordthe product.

Preparative Example 2 Preparation of

A solution of KOH (2.7 g) in ethanol (30.8 g) was magnetically stirredat room temperature. 11-Mercaptoundecanoic acid (5.0 g) was added slowlyto the KOH solution. After the addition was complete, a solution ofiodine (2.9 g) in ethanol (62.2 g) was added and the mixture was stirredfor approximately one hour longer. The mixture was then poured into 1Naqueous HCl and the precipitated solid was isolated by filtration. Thesolid was washed with deionized water and was dried in air to afford 5.0g of product.

Preparative Example 3 Preparation of

A mixture of the carboxy-containing product from Preparative Example 2(2.0 g), thionyl chloride (1.15 g), and methylene chloride (12.6 g) wasstirred and heated at reflux under a nitrogen atmosphere in a roundbottom flask that was fitted with a magnetic stir bar, a refluxcondenser, and a heating mantle. After 6 hours, the mixture was cooledto room temperature and the volatile components were removed using arotary evaporator to afford the product.

Preparative Example 4 Preparation of

The chlorocarbonyl-containing product of Preparative Example 3 (10.9 g)dissolved in methylene chloride (30 mL) was added slowly to amagnetically stirred solution of 2-(2-aminoethoxy)ethanol (9.7 g) andN,N-diisopropylethylamine (6.27 g) in methylene chloride (37 mL) withina round bottom flask. During the addition, the flask was cooled in anice bath. After the addition was complete, the mixture became viscous.An attempt to extract the mixture with deionized water resulted in apartial emulsion. The volatile components were removed using a rotaryevaporator and the remaining mixture was heated to boiling, whichresulted in the precipitation of a white solid. This solid was filteredand was then dissolved in acetonitrile (250 mL). The acetonitrilesolution was stirred and was concentrated by directing a stream ofnitrogen gas onto the surface of the solution. The resultant whitecrystals were isolated by filtration and were dried overnight under astream of nitrogen gas to afford 12.6 g of product.

Preparative Example 5 Preparation of

A mixture of the hydroxy-containing product of Preparative Example 4(3.0 g), succinic anhydride (1.1 g) and triethylamine (1.15 g) washeated within an Erlenmeyer flask for 6 hours. The mixture was allowedto cool to room temperature and methyl alcohol was added to the flask.The product formed a dark thick liquid that was not miscible with themethyl alcohol. The methyl alcohol was decanted away from the darkresidue and the product was then recrystallized from acetonitrile toafford 3.43 g of product.

Preparative Example 6 Preparation of

A mixture of the carboxy-containing product of Preparative Example 5(0.5 g), thionyl chloride (0.15 g), DMF (1 drop) and dichloromethane(2.0 g) was magnetically stirred overnight. The volatile components wereremoved using a rotary evaporator to afford the product (0.52 g).

Preparative Example 7 Preparation of

A mixture of the carboxy-containing product of Preparative Example 2(2.0 g), thionyl chloride (1.15 g), and methylene chloride (12.6 g) washeated under reflux within a round bottom flask that was fitted with areflux condenser, a hose adapter connected to a source of nitrogen gas,and a heating mantle. After 6 hours, the mixture was concentrated todryness using a rotary evaporator. To the flask was then added methylenechloride (12.6 g) and to this solution was added dropwise a mixture ofN-hydroxysuccinimide (1.11 g) and pyridine (0.8 g). The mixture wasstirred at room temperature overnight and then the volatile materialswere removed using a rotary evaporator. The residue was recrystallizedfrom isopropyl alcohol to afford 2.91 g of product.

Preparative Example 8 Preparation of a Multilayer Substrate ofDLG-DLC-polyimide-DLC-DLG

A Model 2480 parallel-plate capacitively coupled reactive ion etcher(obtained from PlasmaTherm, St. Petersburg, Fla.) was used to deposit adiamond-like glass (DLG) coating using a tetramethylsilane plasma onto adiamond-like carbon coating (DLC). The DLC coating was deposited usingan acetylene plasma with the Model 2480 reactive ion etcher onto apolyimide film.

An approximately 20 cm by 30 cm sample of polyimide film (availableunder the trade designation “KAPTON E” from E.I. du Pont de Nemours &Co., Wilmington, Del.) was affixed to the powered electrode of the ionetcher using 3M 811 Adhesive Tape from 3M Company, St. Paul, Minn. Theion etcher chamber was closed and the chamber was pumped to a pressureof 0.67 Pa (0.005 Torr). Oxygen gas was introduced into the chamber at aflow rate of 500 standard cm³ per minute, and the pressure of thechamber was maintained at 6.7 Pa (0.050 Torr). Plasma was ignited andwas sustained at a power of 2000 W for 15 seconds. The oxygen gas flowwas then terminated and the chamber was allowed to pump to a pressure of0.67 Pa (0.005 Torr). Acetylene gas was then introduced into the chamberat a flow rate of 200 standard cm³ per minute, and the pressure of thechamber was maintained at 2 Pa (0.015 Torr). Plasma was ignited and wassustained at a power of 1600 W for 10 seconds. The flow of acetylene gaswas then terminated and the chamber was allowed to pump to a pressure of0.67 Pa (0.005 Torr).

Oxygen gas was again introduced into the chamber at a flow rate of 500standard cm³ per minute and, the pressure of the chamber was maintainedat 20 Pa (0.15 Torr). Plasma was ignited and was sustained at a power of300 W for 10 seconds. With the oxygen gas flow rate maintained at 500standard cm³ per minute, tetramethylsilane gas was introduced into thechamber at a flow rate of 150 standard cm³ per minute. The chamberpressure was maintained at 20 Pa (0.15 Torr). Plasma was ignited and wassustained at a power of 300 W for 12 seconds. The flow oftetramethylsilane gas was terminated. After a period of 1 minute, withboth the flow of oxygen gas and the chamber pressure of 20 Pa (0.15Torr) maintained, plasma was ignited and was sustained at a power of300W for 20 seconds. The flow of oxygen gas was then terminated and thechamber pressure was allowed to pump to a pressure of 0.67 Pa (0.005Torr). The chamber was then opened to the atmosphere and the sample wasremoved from the powered electrode, turned so that the DLG coating facedthe electrode, and was again affixed to the electrode. The sequence ofplasma treatments was repeated to provide a sample of polyimide withsequential layers of DLC and DLG on each side.

Preparative Example 9 Preparation of a Multilayer Substrate ofGlass-DLC-DLG

A 25 mm by 75 mm glass microscope slide (available as “MICRO SLIDESSELECTED” from VWR Scientific, West Chester, Pa.) was treated in aplasma chamber according to the method of Preparative Example 8 tosequentially deposit layers of DLC and DLG onto one side of the glassmicroscope slide.

Preparative Example 10 Preparation of a Multilayer Substrate ofPolyimide-Titanium-Gold

Sequential layers of titanium and gold were deposited by electron beamevaporation onto polyimide film. A 10 cm by 15 cm sample of polyimidefilm (available under the trade designation “KAPTON E” from E. I. DuPont de Nemours & Co., Wilmington, Del.) was affixed to the plate of theplanetary system in a Model Mark 50 high vacuum deposition system(available from CHA Industries, Fremont, Calif.) using metal stationerybinder clips. The chamber was evacuated for approximately 2 hours,during which time the chamber pressure was reduced to approximately6.7×10⁻⁴ Pa (5×10⁻⁶ mm Hg). Titanium metal was deposited at a rate ofapproximately 5 Angstroms per second to a total thickness ofapproximately 200 Angstroms. Deposition of titanium was then terminatedand the system was allowed to cool for approximately 30 minutes. Goldmetal was then deposited onto the titanium layer at a rate ofapproximately 1 Angstrom per second to a total thickness ofapproximately 2000 Angstroms. Deposition of gold was then terminated andthe system was allowed to cool for approximately 30 minutes before thechamber pressure was raised to atmospheric pressure and the samples wereremoved.

Preparative Example 11 Attachment of an N-acyloxysuccinimide ContainingTethering Group to Gold-Coated Silicon Substrate

A 250-micromolar solution of the N-acyloxysuccinimide-containing productof Preparative Example 7 in acetone was prepared. A 1 cm by 1 cm portionof a gold-coated silicon wafer was immersed in the solution for 30minutes, after which time it was removed and was rinsed sequentiallywith ethanol and methanol and was then dried by directing a stream ofnitrogen gas over the treated gold surface for approximately 1 minute.The ellipsometric thickness was determined to be 17 Angstroms and thestatic advancing contact angle of deionized water on the surface wasdetermined to be 50 degrees.

Preparative Example 12 Preparation of Acid Chloride Functionalizedpoly(methylmethacrylate-co-methacrylic acid) Beads

Poly(methylmethacrylate-co-methacrylic acid) beads (available under thetrade designation “MCI GEL CQK31P” from Mitsubishi Chemical Corp.,Tokyo, Japan) (20 g) were combined with cyclohexane (66 g) and thionylchloride (8.3 g) in a round bottom flask fitted with a magnetic stirbar, a reflux condenser and a source of nitrogen gas. The mixture washeated under reflux for 6 hours, during which time nitrogen gas wasslowly passed through the apparatus. The mixture was then allowed tocool to room temperature and filtered. The beads were washed withcyclohexane and were then dried under a stream of nitrogen gas overnightto afford the product.

Preparative Example 13 Preparation of Hydroxyl Functionalizedpoly(methylmethacrylate-co-methacrylic acid) Beads

The acid chloride functionalized poly(methylmethacrylate-co-methacrylicacid) b beads of Preparative Example 12 (20.43 g) were combined with2-(2-aminoethoxy)ethanol (40.0 g) in a round bottom flask fitted with amagnetic stir bar. The mixture was stirred overnight at room temperatureand then the beads were filtered and washed with methanol. The beadswere then dried under a stream of nitrogen gas overnight to afford theproduct.

Example 1 Preparation of

A mixture of dry sodium saccharin (10.25 g), dimethoxyethane (150 mL),and sebacoyl chloride (18.0 g) was stirred overnight at room temperatureunder a nitrogen atmosphere. The mixture was then filtered and filtratewas concentrated using a rotary evaporator. The product residue wastriturated with a 10:1 (v/v) mixture of hexane and toluene and thismixture was cooled and then filtered. The filtered solid was dried toafford 22.1 g of product.

Example 2 Preparation of

A mixture of sodium saccharin dihydrate (1.0 g) and toluene(approximately 15 mL) was magnetically stirred and boiled under refluxin a round bottom flask fitted with a Dean-Stark trap and a refluxcondenser. After 6 hours, the mixture was allowed to cool to roomtemperature and the volatile components were removed using a rotaryevaporator. The entire portion of this material was combined with dryacetone (5.6 g) and succinoyl chloride (2.57 g) in a round bottom flask.The mixture was magnetically stirred at room temperature for 30 minutesafter which time it was filtered. The filtrate was concentrated using arotary evaporator. The residue was then further concentrated byconnecting the flask to a high vacuum pump using a hose adapter. Theflask was heated to 75° C. using an oil bath while the flask wasconnected to the vacuum pump. After approximately 2 hours, the flask wasallowed to cool to room temperature and was disconnected from the pump.The product residue was then triturated with toluene that resulted inthe formation of a solid precipitate. The mixture was filtered and thesolid was dried at room temperature to afford 1.0 g of product.

Example 3 Preparation of

A solution of N-phenyltrifluoromethylsulfonamide (2.2 g) andN,N-diisopropylethylamine (1.3 g) in dry THF (25 mL) is added to astirred solution of 10-undecenoyl chloride (2.0 g) in dry THF (25 mL).The solution is stirred overnight at room temperature and then thevolatile components are removed using a rotary evaporator and then ahigh vacuum pump. A solution of this material is then made in methylenechloride (30 g) in 125 mL screw cap bottle.Platinum(0)-1,3-divinyl-1,1,3,3-tetramethyldisilane complex in xylenesis diluted with methylene chloride to a concentration of approximately1.5 weight percent, and 3 drops of this solution are added to thebottle. The bottle is then sealed and is heated to 60° C. in a waterbath. After 18 hours, the mixture is allowed to cool to room temperatureand additional platinum complex solution (1 drop) is added. The bottleis again sealed and is heated at 60° C. for an additional 24 hours. Themixture is then cooled to room temperature and the volatile componentsare removed using a rotary evaporator to afford the product.

Example 4 Preparation of

A solution of N-phenyltrifluoromethanesulfonamide (0.97 g) andN,N-diisopropylethylamine (0.58 g) in dry THF (3.9 g) was added to astirred solution of the product of Preparative Example 3 (1.0 g) in dryTHF (4.0 g) within a round bottom flask. This mixture was allowed tostir overnight. The volatile components were then removed using a rotaryevaporator. The residue was dissolved in chloroform (40 mL) and waswashed first with 0.1N aqueous HCl (40 mL) and then with saturatedaqueous sodium chloride. The organic phase was dried over magnesiumsulfate and was then filtered. The volatile components were removedusing a rotary evaporator to afford 1.03 g of product.

Example 5 Preparation of

Solid sodium saccharin (0.38 g) was added at room temperature to amagnetically stirred solution of the chlorocarbonyl containing productof Preparative Example 6 (0.75 g) in dry acetone (4.1 g). After mixingovernight, the mixture was poured into deionized water in a beaker andthe resultant solid was filtered and dried overnight in a vacuum oven atroom temperature and 66.7 Pa (0.5 mm Hg) to afford 0.60 g of product.

Example 6 Preparation of

A solution of N-methyltrifluoromethanesulfonamide (0.203 g) andN,N-diisopropylethylamine (0.17 g) in THF (0.8 g) was slowly added to astirred solution of the chlorocarbonyl containing product of PreparativeExample 6 (0.5 g) in THF (2.0 g). The mixture was stirred overnight atroom temperature and then the mixture was poured into deionized waterwithin a beaker. The solid was filtered and was dried overnight in avacuum oven at room temperature and 66.7 Pa (0.5 mm Hg) to afford 0.65 gof product.

Example 7 Preparation of

A solution of N-phenyltrifluoromethanesulfonamide (0.28 g) andN,N-diisopropylethylamine (0.16 g) in THF (1.2 g) was slowly added to astirred solution of the chlorocarbonyl containing product of PreparativeExample 6 (0.5 g) in THF (2.0 g). The mixture was stirred overnight atroom temperature and then the mixture was poured into deionized waterwithin a beaker. The solid was filtered and was dried overnight in avacuum oven at room temperature and 66.7 Pa (0.5 mm Hg) to afford 0.72 gof product.

Example 8 Preparation of

A solution of N-methyltrifluoromethanesulfonamide (0.353 g) andN,N-diisopropylethylamine (0.274 g) in dry THF (1.1 g) was added to astirred solution of the product of Preparative Example 3 (1.0 g) in dryTHF (4.0 g) in a round bottom flask. The mixture was magneticallystirred at room temperature overnight. The mixture was then treated with20 mL of 0.1N aqueous HCl and the mixture was extracted with chloroform.The organic phase was washed with saturated aqueous NaCl and was driedover anhydrous MgSO₄. The volatile materials were removed using a rotaryevaporator. Analysis of the residue by ¹H NMR spectroscopy indicatedthat the reaction was not complete. The product residue was transferredto a round bottom flask and was combined with thionyl chloride (0.3 g)and methylene chloride (5 mL). This mixture was magnetically stirredovernight and then the volatile components were removed using a rotaryevaporator. A solution of N-methyltrifluoromethanesulfonamide (0.353 g)and N,N-diisopropylethylamine (0.274 g) in dry THF (1.1 g) was thenadded to the flask and the mixture was magnetically stirred overnight atroom temperature. The mixture was then treated with 20 mL of 0.1Naqueous HCl and the mixture was extracted with chloroform. The organicphase was washed with saturated aqueous NaCl and was dried overanhydrous MgSO₄. The volatile materials were removed using a rotaryevaporator, and then a high vacuum pump, to afford 0.55 g of product.

Example 9 Preparation of

A mixture of 2,3-dihydro-3-oxobenzisosulfonazole (5.0 g), triethylamine(3.3 g) and acetonitrile (30 g) in a round bottom flask was magneticallystirred under a nitrogen atmosphere and was cooled in an ice bath. Asolution of 10-undecenoyl chloride (6.1 g) in THF (12 g) was slowlyadded to the flask using a pressure-equalizing addition funnel. Themixture was allowed to warm to room temperature and was then filtered.The filtrate was concentrated to dryness using a rotary evaporator andthe residue was triturated with diethyl ether. The resultant solid wasfiltered, washed with diethyl ether, and dried in air at roomtemperature to afford 8.7 g of product.

Example 10 Preparation of

A solution of saccharin (1.8 g) and pyridine (0.8 g) in acetonitrile(8.0 g) was added to a stirred solution of the chlorocarbonyl-containingproduct of Preparative Example 3 (2.2 g) in acetonitrile (7.8 g). Aftermixing overnight, most of the solvent was removed using a rotaryevaporator and the remainder of the mixture was poured into deionizedwater in a beaker. The resultant solid was filtered, washed sequentiallywith isopropyl alcohol and diethyl ether, and was dried overnight in avacuum oven at room temperature and 66.7 Pa (0.5 mm Hg) to afford 3.5 gof product.

Example 11 Preparation of

A mixture of the ethylenically unsaturated containing product of Example9 (4.0 g), trichlorosilane (3.1 g), and methylene chloride (25 g) wascombined in a 125 mL screw cap bottle.Platinum(0)-1,3-divinyl-1,1,3,3-tetramethyldisilane complex in xyleneswas diluted with methylene chloride to a concentration of approximately1.5 weight percent, and 3 drops of this solution were added to thebottle. The bottle was then sealed and was heated to 60° C. in a waterbath. After 18 hours, the mixture was allowed to cool to roomtemperature and additional platinum complex solution (1 drop) was added.The bottle was again sealed and was heated at 60° C. for an additional24 hours. The mixture was then cooled to room temperature and thevolatile components were removed using a rotary evaporator.

Example 12 Attachment of a N-sulfonylaminocarbonyl Containing TetheringGroup to a Gold-Coated Silicon Substrate

A 250-micromolar solution of the disulfide containing product of Example10 in acetone was prepared. A 1 cm by 1 cm portion of a gold-coatedsilicon wafer was immersed in the solution for 30 minutes, after whichtime it was removed and was rinsed sequentially with ethanol andmethanol and was then dried by directing a stream of nitrogen gas overthe treated gold surface for approximately 1 minute. The ellipsometricthickness on the gold side of the multilayer substrate was determined tobe 18 Angstroms. The static advancing contact angle of deionized wateron the surface resulting from the attachment of the tethering groups tothe gold substrate layer was determined to be 62 degrees.

Example 13 Attachment of a N-sulfonylaminocarbonyl Containing TetheringGroup to Multilayer Substrate Of Glass-DLC-DLG

A 1-millimolar solution of the trichlorosilyl containing product ofExample 11 in methylene chloride was prepared. A multilayer substrate ofglass-DLC-DLG, the product of Preparative Example 9, was immersed inthis solution for 30 minutes, after which time it was rinsed withmethylene chloride and was dried by directing a stream of nitrogen gasover the treated gold surface for approximately 1 minute. The staticadvancing contact angle of deionized water on the surface resulting fromthe attachment of tethering groups to the DLG substrate layer wasdetermined to be 63 degrees.

Example 14 Attachment of a N-sulfonylaminocarbonyl Containing TetheringGroup to a Multilayer DLG-DLC-polyimide-DLC-DLG Substrate

A 1 millimolar solution of the trichlorosilyl product of Example 11 inmethylene chloride was prepared. A sample a DLG-DLC-polyimide-DLC-DLGmultilayer substrate, approximately 2.5 cm by 7 cm, the product ofPreparative Example 8, was immersed in this solution for 30 minutes,after which time both sides were rinsed with methylene chloride and thesample was dried by directing a stream of nitrogen gas over both DLGsurfaces for approximately 1 minute each. The static advancing contactangle of deionized water on a surface resulting from attachment of thetethering groups to the DLG substrate layer was determined to be 63degrees.

Example 15 Attachment of a N-sulfonylaminocarbonyl Containing TetheringGroup to a Multilayer Polyimide-Titanium-Gold Substrate

A 1 millimolar solution of the disulfide product of Example 10 inacetone was prepared. A sample a polyimide-titanium-gold multilayersubstrate, approximately 2.5 cm by 7 cm, the product of PreparativeExample 10, was immersed in this solution for 30 minutes, after whichtime both sides were rinsed with acetone and the sample was dried bydirecting a stream of nitrogen gas over the gold surface forapproximately 1 minute each. The static advancing contact angle ofdeionized water on the surface resulting from attachment of thetethering groups to the gold substrate layer was determined to be 63degrees.

Examples 16-17 Immobilization of Lysine with N-sulfonylaminocarbonylContaining Group Attached to a Gold Coated Silicon Substrate

Two 1 cm by 1 cm samples of the product of Example 12 (aN-sulfonylaminocarbonyl containing tethering group attached to agold-coated silicon substrate) were immersed in a 1 millimolar solutionof lysine in carbonate buffer. One of the samples (Example 16) wasremoved from the buffer after 30 minutes and was rinsed with deionizedwater. The ellipsometric thickness was determined as described above.The second sample (Example 17) was removed from the buffer after 90minutes and was rinsed with deionized water before the ellipsometricthickness was determined. The data are given in Table 1 for thethickness of the layer attached to the gold substrate surface.

Comparative Example 1-2 Immobilization of Lysine withN-acyloxysuccinimide-Containing Tethering Group Attached to aGold-Coated Silicon Substrate

Two 1 cm by 1 cm samples of the product of Preparative Example 11(N-acyloxysuccinimide containing tethering group attached to agold-coated silicon substrate) were immersed in a 1 millimolar solutionof lysine in carbonate buffer. One of the samples was removed from thebuffer after 30 minutes (Comparative Example 1) and was rinsed withdeionized water. The ellipsometric thickness was determined as describedabove. The second sample (Comparative Example 2) was removed from thebuffer after 90 minutes and was rinsed with deionized water before theellipsometric thickness was determined. The data are given in Table 1for the thickness of the layer attached to the gold substrate surface.

TABLE 1 Examples 16-17 and Comparative Examples 1-2 Time in Change inLysine Ellipsometric Example Solution Thickness (Angstroms) 16 30 3Comparative 30 2 Example 1 17 90 6 Comparative 90 4 Example 2

Example 18 Immobilization of Fluorescent Labeled IgG with aN-sulfonylaminocarbonyl Containing Tethering Group Attached to aMultilayer Substrate of Glass-DLC-DLG

Fluorescent labeled mouse IgG was reconstituted by mixing 0.55 mL ofdeionized water to give a solution of the IgG with a concentration of 2mg/mL. This solution was diluted with CHES buffer to a IgG concentrationof 50 μg/mL. Successive dilutions were made to give samples with IgGconcentrations of 50 μg/mL, 25 μg/mL, 12.5 μg/mL, and 6.25 μg/mL. Analiquot (15 μL) of each of the IgG solutions was deposited via pipetonto a different portion of the product of Example 13 (aN-sulfonylaminocarbonyl containing tethering group attached to amultilayer substrate of Glass-DLC-DLG). The aliquots were smeared acrossthe surface of the slide using the tip of the pipet. The IgG solutionwas allowed to stand on the glass slide for 30 minutes, then the slidewas washed with sequentially with PBS buffer, PBS buffer containing 0.05weight percent TWEEN 20, and PBS buffer. The slide was then allowed todry in air at room temperature for approximately 1 hour. The slide wasanalyzed using a Model GeneTAC UC-4 scanner (available from GenomicSolutions, Inc., Ann Arbor, Mich.). The results, shown in FIG. 1,indicate that the fluorescent labeled mouse IgG is bound to the surfaceof the substrate. Qualitative fluorescence intensity is highest with themost concentrated fluorescent labeled IgG sample and lowest with theleast concentrated IgG sample.

Example 19 Capture of Staphylococcus aureus with Immobilized IgG onMultilayer Substrate of DLG-DLC-polyimide-DLC-DLG

Rabbit IgG specific to Staphylococcus aureus (rabbit anti Staphylococcusaureus, obtained from Accurate Chemical & Scientific Corp., Westbury,N.Y.) was used at a concentration of 4.52 mg/mL. This solution wasdiluted with CHES buffer to give a solution with a concentration of theIgG of 50 μg/mL. A 1 cm by 1 cm sample of the product of Example 14(N-sulfonylaminocarbonyl containing tethering group attached to amultilayer substrate of DLG-DLC-polyimide-DLC-DLG; that is, thesubstrate was polyimide coated on both sides with a layer of DLC andthen a layer of DLG) was immersed in this solution for 30 minutes afterwhich time it was washed sequentially with PBS buffer, PBS buffercontaining 0.05 weight percent TWEEN 20, and PBS buffer. The sample withimmobilized IgG was then allowed to dry in air at room temperature forapproximately 1 hour.

A solution of acridine orange in deionized water at a concentration of10 mg/mL (obtained from Molecular Probes, Inc., Eugene, Oreg.) wasdiluted to a concentration of 0.1 mg/mL with deionized water. A 500microliter aliquot of this solution was mixed in a centrifuge tube witha 500 microliter aliquot of a suspension of Staphylococcus aureus in PBSbuffer at a concentration of 109 colony forming units per milliliter(cfu/mL). This mixture was allowed to stand at room temperature for 15minutes, after which time it was mixed using a laboratory vortex mixerand was then centrifuged at 8000 rpm. The supernatant liquid was removedusing a pipette and the bacteria were washed three times by adding 500microliters of deionized water to the tube, mixing the contents usingthe vortex mixer, centrifuging the tube at 8000 rpm, and removing thesupernatant liquid. The bacteria were then dispersed in PBS buffer byadding 500 microliters of buffer to the centrifuge tube and mixing thecontents by using the vortex mixer. The concentration of S. aureus inthe buffer was 10⁹ colony forming units per milliliter (10⁹ cfu/mL).

The substrate with immobilized IgG was then affixed to a glassmicroscope slide using double-sided adhesive tape (available from 3MCompany, St. Paul, Minn.) and this construction was immersed in thesuspension of S. aureus in PBS buffer for 1 hour. The sample was thenwashed sequentially with PBS buffer, PBS buffer containing 0.05 weightpercent TWEEN 20, and PBS buffer. The sample was then immersed in a 1weight percent aqueous solution of paraformaldehyde for 15 minutes,after which time it was washed with deionized water. The sample wasanalyzed by confocal microscopy using an Olympus Model FV-300 confocalmicroscope (available from Leeds Precision Inc., Minneapolis, Minn.).The results are shown in FIG. 2.

Comparative Example 3 Exposure of Staphylococcus aureus to MultilayerSubstrate of DLG-DLC-polyimide-DLC-DLG

A 1 cm by 1 cm sample of the substrate of Preparative Example 8(multilayer substrate of DLG-DLC polyimide film-DLC-DLG) was immersedCHES buffer for 30 minutes after which time it was washed sequentiallywith PBS buffer, PBS buffer containing 0.05 weight percent TWEEN 20, andPBS buffer. The substrate was then allowed to dry in air at roomtemperature for approximately 1 hour. The substrate was then immersed ina suspension of Staphylococcus aureus and was then rinsed and immersedin a 1 weight percent aqueous paraformaldehyde solution as described inExample 19. The sample was analyzed by confocal microscopy using anOlympus Model FV-300 confocal microscope (available from Leeds PrecisionInc., Minneapolis, Minn.). The results are shown in FIG. 3.

Examples 20-35 ELISA using a Multilayer Substrate ofDLG-DLC-polyimide-DLC-DLG having Attached N-SulfonylaminocarbonylContaining Tethering Groups

For each of Examples 20-35, a 1 cm by 1 cm sample of the product ofExample 14 (a multilayer substrate of DLG-DLC-polyimide-DLC-DLG withattached N-sulfonylaminocarbonyl containing tethering groups) was placedin a sterile culture tube that contained CHES buffer (1 mL) and variousconcentrations of the antibody anti-human mouse IgG. Four tubes eachcontained a concentration of anti-human mouse IgG in CHES buffer of 5μg/mL, 10 μg/mL, 20 μg/mL, or 50 μg/mL. Each tube was shaken on alaboratory shaker for an exposure time of 5, 10, 30, or 60 minutes. Thebuffer was removed from each tube using a pipette and then the substrateimmobilized IgG sample in each tube was washed three times with PBSbuffer that contained 0.05 weight percent TWEEN 20.

To each tube there was then added 1.5 mL of a solution of 2 weightpercent nonfat dry milk powder (available under the trade designation“NESTLE CARNATION NONFAT DRY MILK POWDER” from Nestle USA, Glendale,Calif.) in PBS buffer. Each tube was placed on the shaker for 1 hourafter which time the solution was removed using a pipette and then thesample in each tube was washed three times with PBS buffer thatcontained 0.05 weight percent TWEEN 20.

A 1 mL aliquot of a solution of biotin-conjugated human IgG in PBSbuffer, at a concentration of 4 μg/mL, was then added to each tube. Thetubes were placed on the shaker for 1 hour after which time the solutionwas removed using a pipette and then the sample in each tube was washedthree times with PBS buffer that contained 0.05 weight percent TWEEN 20.A 1 mL aliquot of a solution of the detecting enzyme SA-HRP in PBSbuffer, at a concentration of 0.5 μg/mL, was added to the tubes. Thetubes were placed on the shaker for 30 minutes, after which time thesolution was removed from each tube using a pipette. Then the samplefilm in each tube was washed three times with PBS buffer that contained0.05 weight percent TWEEN 20.

A 1 mL aliquot of the ABTS indicator solution was added to each tubeand, after 5 minutes, a solution of 1 weight percent aqueous SDSsolution (1 mL) was added. An aliquot of the solution in each tube wastransferred to a standard cuvette and the absorbance of each solution at405 nm was measured using a Model 8453 ultraviolet/visiblespectrophotometer (available from Hewlett-Packard Co., Palo Alto,Calif.) The data are given in Table 2.

TABLE 2 ELISA Examples 20-35 Antibody Absorbance Concentration Exposureat 405 nm Example (μg/mL) Time (min) (relative units) 20 5 5 0.0651 21 510 0.2467 22 5 30 0.3441 23 5 60 0.4367 24 10 5 0.2466 25 10 10 0.415726 10 30 0.4146 27 10 60 0.4418 28 20 5 0.4442 29 20 10 0.4147 30 20 300.4694 31 20 60 0.5261 32 50 5 0.5358 33 50 10 0.4418 34 50 30 0.5082 3550 60 0.5262

Examples 36-41 Immobilization of Lysine to a N-SulfonylaminocarbonylContaining Compound Attached to a Multilayer Substrate of Glass-DLC-DLG

Six samples of the product of Example 13 (a N-sulfonylaminocarbonylcontaining tethering group to attached to a multilayer substrate ofGlass-DLC-DLG) having a static advancing contact angle of deionizedwater of 63 degrees, were immersed in a 30 millimolar solution of lysinein CHES buffer. A sample was removed at time intervals, as shown inTable 3, and was washed with CHES buffer and dried under stream ofnitrogen gas. The contact angle was then measured for the layer attachedto the DLG surface of the multilayer substrate as described above. Thedata are shown in Table 3.

TABLE 3 Examples 36-41 Example Time (min) Contact Angle 36 1 23° 37 223° 38 5 23° 39 10 23° 40 30 23° 41 60 23°

Examples 42-46 Immobilization of HSA to a Gold-Coated Silicon Substratewith N-sulfonylaminocarbonyl Containing Tethering Groups

Five samples of the product of Example 12 (N-sulfonylaminocarbonylcontaining tethering group attached to a gold-coated silicon substrate)were immersed in a 10 micromolar solution of HSA in carbonate buffer atpH 9.6. A sample was removed at time intervals, as shown in Table 4, andwas washed sequentially with deionized water, ethanol, and methanol andwas then measured dried under a stream of nitrogen gas. Theellipsometric thickness was then measured as described above and wascompared to the thickness of the product of Example 12 (18 Angstroms).That is, the thickness of the layer attached to the gold surface of thesubstrate was measured. The data are shown in Table 4.

TABLE 4 Examples 42-46 Increase in Ellipsometric Example Time (min)Thickness (Angstroms) 42 0.1 12 43 15 12 44 30 11 45 60 12 46 90 15

Comparative Examples 4-7 Binding of HSA to aN-acyloxysuccinimide-Containing Tethering Attached to a Gold-CoatedSilicon Substrate

Four samples of the product of Preparative Example 11(N-acyloxysuccinimide containing tethering groups attached to agold-coated silicon substrate) were immersed in a 10 micromolar solutionof HSA in carbonate buffer at pH 9.6. A sample was removed at timeintervals, as shown in Table 5, and was washed sequentially withdeionized water, ethanol and methanol and was then dried under a streamof nitrogen gas. The ellipsometric thickness was then measured asdescribed above and was compared to the thickness of the film asprepared as described in Preparative Example 11 (17 Angstroms). That is,the thickness of the layer attached to the gold surface of the substratewas measured. The data are shown in Table 5.

TABLE 5 Comparative Examples 4-7 Increase in Comparative EllipsometricExample Time (min) Thickness (Angstroms) 4 0.1 2 5 15 5 6 60 5 7 90 10

Example 47 Attachment of a N-methyl-trifluoromethanesulfonamideContaining Tethering Group to a Gold-Coated Silicon Substrate

A 250-micromolar solution of the disulfide containing product of Example8 in methyl ethyl ketone was prepared. A 1 cm by 1 cm portion of agold-coated silicon wafer was immersed in the solution for 30 minutes,after which time it was removed and was rinsed sequentially with ethanoland methanol and was then dried by directing a stream of nitrogen gasover the treated gold surface for approximately 1 minute. Theellipsometric thickness was determined to be 17 Angstroms and the staticadvancing contact angle of deionized water on the surface resulting fromthe attachment of the tethering groups to the gold substrate layer wasdetermined to be 79 degrees.

Example 48 Attachment of a N-phenyl-trifluoromethanesulfonamideContaining Tethering Group to a Gold-Coated Silicon Substrate

A 250-micromolar solution of the disulfide containing product of Example4 in methyl ethyl ketone was prepared. A 1 cm by 1 cm portion of agold-coated silicon wafer was immersed in the solution for 30 minutes,after which time it was removed and was rinsed sequentially with ethanoland methanol and was then dried by directing a stream of nitrogen gasover the treated gold surface for approximately 1 minute. Theellipsometric thickness was determined to be 23 Angstroms and the staticadvancing contact angle of deionized water on the surface resulting fromthe attachment of the tethering groups to the gold substrate layer wasdetermined to be 73 degrees.

Example 49 Immobilization of 1-aminododecane withN-methyltrifluoromethanesulfonamine-Containing Group Attached to a GoldCoated Silicon Substrate

A 1 cm by 1 cm sample of the product of Example 47 (aN-methyltrifluoromethanesulfonamide containing tethering group attachedto a gold-coated silicon substrate) was immersed in a 1 millimolarsolution of 1-aminododecane in ethanol. The sample was removed from thesolution after 2 hours and was rinsed sequentially with ethanol andmethanol and was then dried under a stream of nitrogen gas. Theellipsometric thickness was determined to be 21 Angstroms and the staticadvancing contact angle of deionized water on the surface was determinedto be 86 degrees.

Example 50 Immobilization of didodecylamine withN-methyltrifluoromethanesulfonamine-Containing Group Attached to a GoldCoated Silicon Substrate

A 1 cm by 1 cm sample of the product of Example 47 (aN-methyltrifluromethanesulfonamide containing tethering group attachedto a gold-coated silicon substrate) was immersed in a 1 millimolarsolution of didodecylamine in ethanol. The sample was removed from thesolution after 2 hours and was rinsed sequentially with ethanol andmethanol and was then dried under a stream of nitrogen gas. Theellipsometric thickness was determined to be 19 Angstroms and the staticadvancing contact angle of deionized water on the surface was determinedto be 78 degrees.

Example 51 Immobilization of 1-aminododecane withN-phenyltrifluoromethanesulfonamine-Containing Group Attached to a GoldCoated Silicon Substrate

A 1 cm by 1 cm sample of the product of Example 48 (aN-phenyltrifluromethanesulfonamide containing tethering group attachedto a gold-coated silicon substrate) was immersed in a 1 millimolarsolution of 1-aminododecane in ethanol. The sample was removed from thesolution after 2 hours and was rinsed sequentially with ethanol andmethanol and was then dried under a stream of nitrogen gas. Theellipsometric thickness was determined to be 26 Angstroms and the staticadvancing contact angle of deionized water on the surface was determinedto be 78 degrees.

Example 52 Immobilization of Didodecylamine withN-phenyltrifluoromethanesulfonamine-Containing Group Attached to a GoldCoated Silicon Substrate

A 1 cm by 1 cm sample of the product of Example 48 (aN-phenyltrifluromethanesulfonamide containing tethering group attachedto a gold-coated silicon substrate) was immersed in a 1 millimolarsolution of didodecylamine in ethanol. The sample was removed from thesolution after 2 hours and was rinsed sequentially with ethanol andmethanol and was then dried under a stream of nitrogen gas. Theellipsometric thickness was determined to be 23 Angstroms and the staticadvancing contact angle of deionized water on the surface was determinedto be 78 degrees.

Example 53 Capture of Staphylococcus aureus With immobilized IgG onMultilayer Substrate of Polyimide-Titanium-Gold

Rabbit IgG specific to Staphylococcus aureus (rabbit anti Staphylococcusaureus, obtained from Accurate Chemical & Scientific Corp., Westbury,N.Y.) was used at a concentration of 4.52 mg/mL. This solution wasdiluted with CHES buffer to give a solution with a concentration of theIgG of 50 μg/mL. A 1 cm by 1 cm sample of the product of Example 15(N-sulfonylaminocarbonyl containing tethering group attached to amultilayer substrate of polyimide-titanium-gold as described inPreparative Example 10) was immersed in this solution for 30 minutesafter which time it was washed sequentially with PBS buffer, PBS buffercontaining 0.05 weight percent TWEEN 20, and PBS buffer. The sample withimmobilized IgG was then allowed to dry in air at room temperature forapproximately 1 hour.

A solution of acridine orange in deionized water at a concentration of10 mg/mL (obtained from Molecular Probes, Inc., Eugene, Oreg.) wasdiluted to a concentration of 0.1 mg/mL with deionized water. A 500microliter aliquot of this solution was mixed in a centrifuge tube witha 500 microliter aliquot of a suspension of Staphylococcus aureus indeionized water at a concentration of 10⁹ colony forming units permilliliter (cfu/mL). This mixture was allowed to stand at roomtemperature for 15 minutes, after which time it was mixed using alaboratory vortex mixer and was then centrifuged at 8000 rpm. Thesupernatant liquid was removed using a pipette and the bacteria werewashed three times by adding 500 microliters of deionized water to thetube, mixing the contents using the vortex mixer, centrifuging the tubeat 8000 rpm, and removing the supernatant liquid. The bacteria were thendispersed in PBS buffer by adding 500 microliters of buffer to thecentrifuge tube and mixing the contents by using the vortex mixer. Theconcentration of S. aureus in the buffer was 109 colony forming unitsper milliliter (10⁹ cfu/mL).

The substrate with immobilized IgG was then affixed to a glassmicroscope slide using double-sided adhesive tape (available from 3MCompany, St. Paul, Minn.) and this construction was immersed in thesuspension of S. aureus in PBS buffer for 1 hour. The sample was thenwashed sequentially with PBS buffer, PBS buffer containing 0.05 weightpercent TWEEN 20, and PBS buffer. The sample was then immersed in a 1weight percent aqueous solution of paraformaldehyde for 15 minutes,after which time it was washed with deionized water. The sample wasanalyzed by confocal microscopy using an Olympus Model FV-300 confocalmicroscope (available from Leeds Precision Inc., Minneapolis, Minn.).The results are shown in FIG. 4.

Comparative Example 8 Exposure of Staphylococcus aureus to MultilayerSubstrate of Polyimide-Titanium-Gold

A 1 cm by 1 cm sample of the substrate of Preparative Example 10(multilayer substrate of polyimide film-titanium-gold) was immersed CHESbuffer for 30 minutes after which time it was washed sequentially withPBS buffer, PBS buffer containing 0.05 weight percent TWEEN 20, and PBSbuffer. The substrate was then allowed to dry in air at room temperaturefor approximately 1 hour. The substrate was then immersed in asuspension of S. aureus and was then rinsed and immersed in a 1 weightpercent aqueous paraformaldehyde solution as described in Example 53.The sample was analyzed by confocal microscopy using an Olympus ModelFV-300 confocal microscope (available from Leeds Precision Inc.,Minneapolis, Minn.). The results are shown in FIG. 5.

Example 54 Attachment of a N-sulfonylaminocarbonyl Containing TetheringGroup to a Substrate of poly(methylmethacrylate-co-methacrylic Acid)Beads

The hydroxyl functionalized beads of Preparative Example 13 (2.0 g) werecombined with NMP (15 mL) in a round bottom flask that was fitted with amagnetic stir bar. The acid chloride product of Example 2 (0.19 g) inNMP (5 mL) was added to the flask. Ethyldiisopropyl amine (0.09 g) inNMP (5 mL) was then added to the flask and the mixture was magneticallystirred at room temperature overnight. The beads were then filtered,washed with isopropyl alcohol and were dried under a stream of nitrogengas to afford the product as white beads.

Example 55 Immobilization of FITC-Albumin with a N-sulfonylaminocarbonylContaining Tethering Group Attached topoly(methylmethacrylate-co-methacrylic Acid) Beads

The poly(methylmethacrylate-co-methacrylic acid) beads with theN-sulfonylaminocarbonyl containing tethering group of Example 54 (50 mg)was combined with a solution of FITC-albumin (1 mL) in a centrifugetube. The tube was placed on a laboratory rocker for 45 minutes. Thebeads were then washed by centrifuging the tube, decanting thesupernatant liquid and then adding PBS buffer having a pH of 7.2 (11mL), again centrifuging the tube and again decanting the supernatantliquid. This washing with PBS buffer was repeated for a total of fourwashing cycles to afford beads that were yellow-orange in color. Thecolor of the beads thus obtained was compared to the color of the beadsof Example 54 that were not treated with FITC-albumin, which were whitein color.

1. An article comprising: a substrate; a substrate-attached tetheringgroup comprising a reaction product of a complementary functional groupG on a surface of the substrate with a compound of Formula I

 wherein X¹ is a substrate-reactive functional group selected from acarboxy, halocarbonyl, halocarbonyloxy, cyano, hydroxy, mercapto,isocyanato, halosilyl, alkoxysilyl, acyloxysilyl, azido, aziridinyl,haloalkyl, tertiary amino, primary aromatic amino, secondary aromaticamino, disulfide, alkyl disulfide, benzotriazolyl, phosphono,phosphoroamido, or phosphato; Y¹ is a single bond or a divalent groupselected from an alkylene, heteroalkylene, arylene, carbonyl,carbonyloxy, carbonylimino, oxy, thio, —NR^(d)—where R^(d) is hydrogenor alkyl, or combinations thereof; Z¹is (CO)R^(a) wherein R^(a) togetherwith R¹ and groups to which they are attached form a four to eightmembered heterocyclic or heterobicyclic group having a nitrogenheteroatom and a sulfur heteroatom, wherein said heterocyclic orheterobicyclic group can be fused to an optional aromatic group,optional saturated or unsaturated cyclic group, or optional saturated orunsaturated bicyclic group; r is equal to 1 when X¹ is a monovalentgroup or equal to 2 when X¹ is a divalent group; G is the complementaryfunctional group capable of reacting with X¹ to form an ionic bond,covalent bond, or combinations thereof, wherein the group G is selectedfrom hydroxy, mercapto, primary aromatic amino group, secondary aromaticamino group, secondary aliphatic amino group, azide, carboxyl,carboxylic acid anhydride, isocyanate, halocarbonyl, halocarbonyloxy,acrylate, silanol, or nitrile; and said tethering group is unsubstitutedor substituted with a halo, alkyl, alkoxy, or combinations thereof. 2.The article of claim 1, wherein the compound is of formula

where X¹ is monovalent or

where X¹ is divalent.
 3. The article of claim 1, wherein the substratecomprises a polymeric material.
 4. The article of claim 1, wherein thesubstrate comprises a polyimide or polyester film.
 5. The article ofclaim 1, wherein the substrate is multilayered and has an outer layercomprising gold.
 6. The article of claim 1, wherein the substrate ismultilayered and has an outer layer comprising diamond-like glass. 7.The article of claim 1, wherein the substrate is a multilayer substratecomprising; a support layer comprising polyimide or polyester; an outerlayer comprising diamond-like glass; and a layer of diamond-like glasspositioned between the support layer and the outer layer.
 8. The articleof claim 1, wherein the substrate is in the form of a bead.